Sensory and autonomic innervation of the facet joint in the rat lumbar spine

The presence of sensory and autonomic nerves in the synovial membrane of the lumbar facet joint in rats was investigated by immunohistochemistry. Substance P and calcitonin gene-related peptide immunoreactivities, representing sensory nerves, were observed as varicose fibers in the synoviocyte layer. The fibers were predominantly nonvascular. The autonomic innervation was identified by the presence of neuropeptide Y- and tyrosine hydroxylase-positive fibers. Most of these fibers were found adjacent to or within blood vessel walls. Immunoreactivity to vasoactive intestinal polypeptide was seen in varicose nerve terminals in the synoviocyte layer, mostly unrelated to blood vessels. There is accumulating evidence of an involvement of both the sensory and sympathetic nervous systems in inflammatory joint disease. The neuropeptides now identified in lumbar facet joints may prove to play a significant role in the pathogenesis of low-back pain.     

A role for HEM2 in cadmium tolerance

The innervation of the knee joint synovial membrane of the guinea pig, i.e., the synoviocyte layer, the subjacent connective tissue and the connective tissue region beneath, was analyzed with immunohistofluorescence and electron microscopy. A screening of the innervation with antibodies against the general axon marker -- protein gene product (PGP) 9,5 -- revealed the presence of nerve fibers distributed in various regions of the knee joint synovial membrane. Confirming previous studies, some of these nerve fibers stained with antibodies to tyrosine hydroxylase (TH), neuropeptide Y (NPY), substance P (SP), calcitonin gene-related peptide (CGRP), and vasoactive intestinal polypeptide (VIP). In addition, dynorphin (DYN)-containing fibers were detected, which have not been reported previously in normal joints. In general, the immunoreactive fibers were observed close to the synoviocytes and at blood vessels. Fibers with colocalization of NPY- and TH-like immunoreactivities (LIs), as well as of DYN- and TH-LIs were demonstrated. In the electron microscope, bundles of unmyelinated fibers as well as single fibers were found in the connective tissue region below the synoviocytes. Varicose parts of the nerve fibers contained mainly small, clear vesicles. Small and large dense-cored vesicles were also seen, but less frequently. Denser portions of the plasma membranes of some axons were observed in these regions, facing the extracellular space. Myelinated fibers were also observed in some nerve bundles. These findings emphasize the complex innervation of the synovial membrane, with nerve fibers containing a host of neuroactive substances. Altogether, these fibers are probably involved in many functions such as vasoregulation and control of synovial secretion in addition to being a source of mediators in joint inflammation.     

Overexpression of nerve growth factor in skin causes preferential increases among innervation to specific sensory targets

The impact of increased levels of skin-derived nerve growth factor (NGF) neurotrophin on sensory and sympathetic innervation to the mouse mystacial pad and postero-orbital vibrissae was determined. Consistent with an approximate doubling of neuron number in trigeminal and superior cervical ganglia, many components of the sensory and sympathetic innervation were substantially enhanced. Although the increased number of neurons raised the possibility that all types of innervation were increased, immunohistochemical analysis indicated that enhanced NGF production had a differential effect upon sensory innervation, primarily increasing unmyelinated innervation. This increased innervation occurred in specific locations known to be innervated by small, unmyelinated fibers, suggesting that NGF modulated sensory innervation density, but not targeting. In contrast, sympathetic innervation was not only increased but also was distributed to some aberrant locations. In the intervibrissal fur of the mystacial pad, both the number of sensory axons and branches appeared increased, whereas in vibrissal follicle sinus complexes, only branching increased. In some areas, sensory ending density was lower than expected based upon the size of the source nerve bundles suggesting that many axons and branches were surviving but failing to form functional endings. Furthermore, the immunochemical profile of innervation was altered in some sensory populations as demonstrated by the coexistence of RT-97 neurofilament labeling in calcitonin gene-related peptide (CGRP) positive axons, by the loss of substance P colocalization in some CGRP axons, and by an absence of neuropeptide Y labeling in tyrosine hydroxylase positive sympathetic axons. Collectively, these results indicate that the NGF mediated increase in neuron number may be selective for particular sets of innervation and that increases among some populations may result from phenotypic switching.     

Permanent outdoor housing for woodchucks, Marmota monax

Results of investigations on the occurrence of nerve fibres and endings in the synovial membrane of the knee and elbow joint in the cat are reported. The stratum synoviale contains only autonomic fibres, running in the adventitia of arteries. Free nerve endings are lacking in the stratum synoviale. Simple Pacinian corpuscles with an inner core are occasionally observed in the border zone between the stratum synoviale and fibrosum. The ultrastructure of these endorgans resembles that of Pacinian corpuscles in the hairless and hairy skin of the cat.     

Distribution of SP- and CGRP-immunoreactivity in the cat's larynx

The distribution of substance P (SP) and calcitonin gene-related peptide (CGRP) immunoreactive (ir) fibres in the cat's larynx was investigated utilizing immunohistochemistry. Many SP- and CGRP-ir fibres with varicosities were found within and below the epithelium and along the basement membrane of the mucosa of all different regions except in the membranous portion of the vocal fold. In the subepithelium, some SP- and CGRP-ir nerve bundles and nerve fibres were recognized around the vessels and glands. In the mucosa, the pattern of distribution and the density of SR-ir fibres were similar to those of CGRP-ir fibres. These reactive fibres were denser in the supraglottic region than in the subglottic region. In the taste bud-like structures, only SP-ir fibres appeared, whereas in the motor endplates, CGRP-reaction was found exclusively. The present findings suggest that the regional distribution of SP- and CGRP-immunoreactivity might be related with sensory and autonomic innervation in the larynx.     

Dental innervation and CGRP in adult p75-deficient mice

Adult dental tissues have unusual neurotrophin biology. Pulpal fibroblasts express nerve growth factor (NGF) and the low-affinity p75 neurotrophin receptor, their sensory nerve fibers express p75 and trk A, and pulpal sympathetic fibers lack p75. Following tooth injury, there is increased pulpal NGF, sprouting of sensory nerve endings, and increased immunoreactivity for the sensory neuropeptide calcitonin gene-related peptide (CGRP). In the present study, we have analyzed tooth structure and innervation of pulp and periodontal ligament in young (6-8 weeks, 3 months) and older (5-12 months) adult mice carrying a null mutation in the p75 gene and compared the results with those of age-matched wild-type controls. Our hypotheses were that tooth structure would be abnormal and that pulpal innervation would be greatly reduced because it consists primarily of nociceptive fibers that have been found to be severely depleted in skin of p75(-/-) mice. Tissues were fixed, X-rayed for gross dental morphology, decalcified, and analyzed for immunoreactivity for CGRP and for a general nerve marker, protein gene product 9.5. Radiographs showed worn-down molar crowns in p75-deficient mice. Light microscopy confirmed the accelerated molar wear and showed intense CGRP immunoreactivity in pulp nerve endings of mutant mice, compared with a gradual decrease in CGRP intensity in controls during normal aging. The CGRP intensity in 5-12-month-old pairs of mice was threefold greater in the mutants (P < 0.03), and in younger mice the mutant always had more CGRP than its matched control. The innervation of molar ligament in all p75-deficient mice was similar to that of controls except there was nerve sprouting near bone loss in mutants. The incisors of mutant mice did not have unusual wear and their pulpal CGRP immunoreactivity remained normal, but their periodontal ligament had fewer thin branched nerve endings at all ages. Thus, most innervation of teeth and their supporting tissues developed normally, and the only neural changes in p75(-/-) mutant mice were the reduction of incisor ligament sensory receptors and increased molar CGRP. Sensory nerves in teeth gradually lose neuropeptide intensity during aging, but that did not happen in the mutant mice, suggesting that the accelerated molar wear stimulated persistent high levels of CGRP.     

Innervation of cholinergic vestibular efferent system in vestibular periphery of rats

The innervation of cholinergic efferent fibers in the vestibular endorgans of the rats was investigated using a modified preembedding immunostaining technique of immunoelectron microscopy. A monoclonal antibody to choline acetyltransferase (ChAT) was used as a marker of cholinergic fibers. It was found that there were four types of cholinergic innervation in the vestibular endorgans of the rat: (1) cholinergic nerve endings formed axo-dendritic synapses with afferent chalice surrounding the type I sensory hair cells; (2) cholinergic nerve endings formed axo-somatic synapses with type II hair cells; (3) cholinergic fibers synapse with afferent nerve fibers and (4) a synaptic contact developed between cholinergic nerve endings. The results demonstrated that a multiform innervation of the cholinergic efferents exists in the rats vestibular periphery.     

Neural emergency system in the stomach

The maintenance of gastric mucosal integrity depends on the rapid alarm of protective mechanisms in the face of pending injury. Afferent neurons of extrinsic origin constitute an emergency system that is called into operation when the gastric mucosa is endangered by acid and other noxious chemicals. The function of these chemoceptive afferents can be manipulated selectively and explored with the excitotoxin capsaicin. Most of the homeostatic actions of capsaicin-sensitive afferents are brought about by peptides released from their peripheral endings in the gastric wall. When stimulated, chemoceptive afferents enhance gastric blood flow and activate hyperemia-dependent and hyperemia-independent mechanisms of protection and repair. In the rodent stomach, these local regulatory roles of sensory neurons are mediated by calcitonin gene-related peptide acting via calcitonin gene-related peptide 1 receptors and neurokinin A acting via neurokinin 2 receptors, with both peptides using nitric oxide as their common messenger. In addition, capsaicin-sensitive neurons form the afferent arc of autonomic reflexes that control secretory and motor functions of the stomach. The pathophysiological potential of the neural emergency system is best portrayed by the gastric hyperemic response to acid backdiffusion, which is signaled by afferent nerve fibers. This mechanism limits damage to the surface of the mucosa and creates favorable conditions for rapid restitution and healing of the wounded mucosa.     

Modified MACOP-B chemotherapy for intermediate and high grade non Hodgkins lymphomas

The distribution of calcitonin gene-related peptidergic (CGRP) nerve endings in rat nasal mucosa was investigated with immunocytochemical technique (ABC method). The results showed that CGRP endings had a robust localization along the walls of arterioles, venules and around the acini of glands, with the endings near the wall of arterioles being most strongly stained. The typical morphology of CGRP endings could be recognized as multistage-branched terminals with rosary varicosities. CGRP immunoreactive neurons were also found in the trigeminal ganglion (TG). Supported by morphological evidence, we suggest that CGRP endings in rat nasal mucosa are the peripheral fibers originating from the ganglion cells in TG. These fibers may be involved in the perception of mucosal stimuli and the propagation of nerve impulse to the central terminals and will thus release CGRP in the brain stem. There is also possibility that CGRP may be peripherally released following local stimulation in the nasal mucosa and exert influences on the mucosal functions.     

Innervation of the lumbar facet joints. Origins and functions

STUDY DESIGN: The levels of dorsal root ganglia and paravertebral sympathetic ganglia innervating the lumbar facet joint were investigated in rats using the retrograde transport method. The pathways and functions of the nerve fibers supplying the lumbar facet joint were determined immunohistochemically. OBJECTIVES: To study lumbar facet pain in relation to its innervation. SUMMARY OF BACKGROUND DATA: The lumbar facet joints have been reported to be innervated segmentally. Little is known, however, about the origins and functions of the nerve fibers. METHODS: Cholera toxin B subunit, a neural tracer, was placed in the L5-L6 facet joint, and the bilateral dorsal root ganglia and paravertebral sympathetic ganglia were examined immunohistochemically. The serial sections of lumbar vertebrae of newborn rats and the sections of the facet joint capsules, dorsal root ganglia, and paravertebral sympathetic ganglia of adult rats were investigated immunohistochemically. The pathways of the nerve fibers supplying the facet joint were reconstituted. RESULTS: Labeled neurons existed in ipsilateral dorsal root ganglia from L1 to L5 and in paravertebral sympathetic ganglia from T12 to L6. The dorsal ramus of the spinal nerve and rami communicantes were connected to each other by calcitonin gene-related peptide immunoreactive fibers and dopamine beta-hydroxylase immunoreactive fibers. CONCLUSIONS: The L5-L6 facet joint was innervated by ipsilateral dorsal root ganglia and paravertebral sympathetic ganglia, segmentally and nonsegmentally. Some of the sensory fibers from the facet joint may pass through the paravertebral sympathetic trunk, reaching L1 and/or L2 dorsal root ganglia. Inguinal and/or anterior thigh pain with lower lumbar facet joint lesions may be explained as referred pain.     

The medial capsule of the human temporomandibular joint

PURPOSE: Attachments of the medial capsule of the temporomandibular joint (TMJ) to structures other than the medial fossa wall are thought to exist and to have functional significance. This study evaluated these relationships. MATERIALS AND METHODS: The anatomic relationships between the medial capsule and other medial structures, the sphenomandibular ligament, discomalleolar ligament, and auriculotemporal nerve, were examined in 14 cadaver heads. RESULTS: The results showed that the sphenomandibular ligament attaches separately from the medial capsule of the TMJ and therefore has no functional significance to the biomechanics of the joint. The discomalleolar ligament was found to be a continuation of the retrodiscal tissues and minimally associated with the medial capsule. The auriculotemporal nerve was not found to be in a relationship with the medial aspect of the condyle to the extent that mechanical irritation is possible during TMJ movement or disc displacement.     

Behavioural topography in the striatum: differential effects of quinpirole and D-amphetamine microinjections

The structure of the nervous network and the distribution of tyrosine hydroxylase (TH)- and various neuropeptide-containing nerves were immunohistochemically studied in the glottis of the dog. The nervous network in the glottis revealed apparent regional differences in morphology. The nervous network in the cartilaginous vocal fold of the posterior glottis consisted of nerve bundles running parallel to the edge of the vocal fold. Only a small number of nerve bundles were observed in the anterior glottis, specifically in membranous vocal fold. In the subepithelial layer of the posterior glottis, a moderate number of galanin (GAL)-immunoreactive nerve fibers were observed, while only a few fibers were present in the anterior glottis. Numerous vasoactive intestinal peptide (VIP)-, GAL-, methionine-enkephalin (ENK)- and TH-immunoreactive nerve fibers were observed within and around the laryngeal submucosal seromucous gland. Many TH- and neuropeptide Y (NPY)-immunoreactive fibers were arranged around the blood vessels. In the epithelia, free nerve endings with immunoreactivity for substance P (SP) and calcitonin gene-related peptide (CGRP) was observed. Furthermore, nerve cell bodies with SP-, VIP-, GAL-, ENK-, and NPY-immunoreactivity were observed in the deep region of the submucosal layer. The results from the present study suggest that there is autonomic regulation of the glottis. Regional structural differences in the nervous network of the glottis may reflect functional differences.     

Ultrastructural study on adhesions in internal derangement of the temporomandibular joint

PURPOSE: This study examined the ultrastructural characteristics of adhesions in the upper joint compartment of temporomandibular joint (TMJ). MATERIALS AND METHODS: Tissue biopsy specimens of adhesions were obtained during arthroscopic operation on 36 joints in 22 patients with internal derangement (ID). The biopsy specimens were examined by light and transmission electron microscopy. RESULTS: Adhesions were grossly divided into two types based on arthroscopic observation: 1) a band-like type, which connected the articular fossa and TMJ disc, and 2) a pseudowall-like type, which faced the synovial fluid and was lined by articular tissue. Two types of collagen arrangement were observed at the electron microscopic level: orderly arranged collagen bundles and randomly arranged collagen bundles. Orderly arranged collagen bundles were prominent in the band-like adhesions. In pseudowall-like adhesions, mainly the randomly arranged collagen bundles were seen. However, in some dense fiber parts, orderly arranged collagen bundles also were observed. In other pseudowall-like adhesions, only orderly arranged collagen bundles were seen. Elastic fibers were abundant in some pseudowall-like adhesions with randomly arranged collagen bundles. There were no elastic fibers in the band-like adhesions, some dense fiber parts of the pseudowall-like adhesion, pseudowall-like adhesions consisting of only orderly arranged collagen bundles, and in the synovial membrane. CONCLUSION: The different arrangement of collagen fibers and presence or absence of elastic fibers were observed in the two types of adhesions. These findings served to show that extracellular components correspond to a dysfunction involving an ID of TMJ.     

Nerve endings with structural characteristics of mechanoreceptors in the human scleral spur

PURPOSE: The innervation of the scleral spur region was investigated to learn whether mechano-receptors are present in this region. METHODS: Serial tangential sections and whole-mount preparations of the scleral spur region of 18 human eyes of different ages were investigated with electronmicroscopic and immunohistochemical methods. For immunohistochemistry antibodies against neurofilament-proteins, synaptophysin, substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), tyrosine-hydroxylase, dopamine-beta-hydroxylase, and acetylcholinesterase were used. RESULTS: Club- or bulb-shaped nerve endings with a diameter of 5 microns to 25 microns were identified in the scleral spur region throughout the whole circumference of the eyes. The terminals derive from myelinated axons with a diameter of approximately 3 microns and stain with antibodies against neurofilament-proteins and synaptophysin but do not stain for tyrosine-hydroxylase, dopamine-beta-hydroxylase, acetylcholinesterase, NPY, VIP, SP, or CGRP. Electronmicroscopically, the endings contain abundant neurofilaments, granular and agranular vesicles of different sizes, numerous mitochondria, and lysosome-like lamellated structures. The endings are incompletely ensheathed by Schwann cells. Those areas of the cell membrane of the endings that are not covered by Schwann cells are in intimate contact with the fibrillar connective tissue elements of the scleral spur. CONCLUSION: These structural features are highly characteristic for mechanoreceptive nerve endings in other tissues of the human body. The authors therefore hypothesize that the club-or bulb-shaped nerve endings in the human scleral spur are afferent mechanoreceptors that measure stress or strain in the connective tissue elements of the scleral spur. Such changes might be induced by ciliary muscle contraction and/or by changes in intraocular pressure.     

Morphologic changes in the elastic fibers of the temporomandibular joint after experimental disc perforation in the rabbit

PURPOSE: Elastic fibers in the attachment regions of the temporomandibular joint (TMJ) are considered important in the movements of the disc during jaw motion. This study was designed to determine whether there are change in the elastic fibers of the TMJ when the disc is perforated for a long period. MATERIALS AND METHODS: Each TMJ of five rabbits was surgically exposed, and a mediolateral perforation was made in the center of the articulating region of the disc with a scalpel. Five untreated and three sham-operated rabbits were used as controls. The TMJs were removed after 10 months and processed for histochemical demonstration of elastic fibers. RESULTS: Osteoarthritic changes and a significant decrease in number of the elastic fibers in the anterior-inferior attachment and posterior-inferior attachment of the TMJ disc were observed after disc perforation. Conversely, elastic fibers appeared in the fibrous tissue on the resorbed bone of the articular eminence. CONCLUSIONS: Experimental disc perforation changes the distribution and density of the elastic fiber in the TMJ as well as causes osteoarthritis. A knowledge of these facts may improve our understanding of perforation of the disc.     

Immunocytochemical co-localization of substance P and calcitonin gene-related peptide in afferent renal nerve soma of the rat

Substance P, calcitonin gene-related peptide and somatostatin immunoreactivities have been demonstrated in putative afferent renal nerve fibers in the rat. Utilizing retrograde-tracing and immunohistochemistry, we labeled afferent renal nerve soma throughout dorsal root ganglia T9 to L1. Most (85%) of afferent renal nerve perikarya were immunoreactive for calcitonin gene-related peptide, 21% had substance P immunoreactivity and none had somatostatin immunoreactivity. All renal afferents immunoreactive for substance P also contained calcitonin gene-related peptide. These results provide evidence that calcitonin gene-related peptide and substance P are present and co-localized in afferent renal nerves, and therefore, mediate transmission of afferent renal input to the spinal cord in the rat.     

Calcitonin gene-related peptide- and substance P-like-immunoreactive innervation of the anterior pituitary in the rat

In our previous studies substantial amounts of substance P- and calcitonin gene-related peptide-like-immunoreactive nerve fibers have been identified in the anterior pituitary of the monkey and the dog. They were found to be in close proximity to the gland cells, even making synaptic contacts with some types of the gland cells. The present study investigated in detail the calcitonin gene-related peptide- and substance P-like immunoreactivities of the anterior pituitary in the rat. Though the immunoreactive fibers were not as abundant as in the anterior pituitary of the monkey and the dog, they still appeared in notable amounts. The calcitonin gene-related peptide- and substance P-like-immunoreactive nerve fibers occurred mostly as thin, tortuous, and densely varicose fibers, weaving among the gland cells. They are widely distributed, more in the central part of the gland. Double-immunostaining proved nearly complete co-localization of these two peptides in the nerve fibers. It is hypothesized that the anterior pituitary can be regulated by direct neural factors as well as humoral factors.     

Distribution and origin of peptide-containing nerve fibres in the rat and human mammary gland

The structures in the mammary gland involved in milk ejection have been investigated with regard to their relation to different types of peptidergic nerve fibres and their origin. Lactating rats were studied with immunohistochemistry focusing on the nipple, the parenchyma, the mammary blood vessels and the mammary nerve. The human mammary gland was also analysed. In the mammary gland from rat and human, nerve endings in the subepidermis, around smooth muscle cells in the nipple, in the connective tissue surrounding lactiferous ducts and alveoli in the nipple and in the parenchyma of the mammary gland showed immunoreactivity for calcitonin gene-related peptide, substance P, vasoactive intestinal polypeptide, peptide histidine isoleucine, neuropeptide Y, galanin and tyrosine hydroxylase, whereas dynorphin-positive nerve fibres could not be detected. The mammary nerve contained calcitonin gene-related peptide, vasoactive intestinal polypeptide, neuropeptide Y and tyrosine hydroxylase immunoreactivities; the adventitia of the mammary artery contained nerve fibres immunoreactive for neuropeptide Y and tyrosine hydroxylase, while vasoactive intestinal polypeptide-, peptide histidine isoleucine-, calcitonin gene-related peptide- and substance P-positive fibres were found in the tissue surrounding the artery. The wall of the mammary vein had nerve terminals immunoreactive for neuropeptide Y, tyrosine hydroxylase, calcitonin gene-related peptide and substance P. With the help of retrograde tracing using wheat germ agglutinin in combination with immunohistochemistry, projections of calcitonin gene-related peptide-immunoreactive cells in the dorsal root ganglia to the nipple were established. Neurons in the sympathetic stellate ganglion containing neuropeptide Y and tyrosine hydroxylase also projected to the mammary gland. Moreover retrogradely-labelled cells were found in the nodose ganglion, and they were vasoactive intestinal polypeptide-immunoreactive. These results demonstrate a rich distribution of different types of nerve fibres in structures of the mammary gland related to milk ejection. These nerve fibres and their peptides may be involved in the local control of milk ejection.     

Vagal and spinal afferent innervation of the rat esophagus: a combined retrograde tracing and immunocytochemical study with special emphasis on calcium-binding proteins

Vagal afferent neurons contain a variety of neurochemical markers and neuroactive substances, most of which are present also in dorsal root ganglion cells. To test for the suitability of the calcium-binding protein calretinin as a specific marker for vagal afferent fibers in the periphery, immunocytochemistry for this protein was combined with retrograde tracing. Nerve fibers in the rat esophagus, as well as vagal and spinal sensory neurons innervating the esophagus, were investigated for co-localization of calretinin with calbindin, calcitonin gene-related peptide, and NADPH diaphorase. The results indicated that calretinin immunocytochemistry demonstrates neuronal structures known as vagal afferent from other studies, in particular intraganglionic laminar endings. A few enteric neurons whose distribution was unrelated to intraganglionic laminar endings also stained for calretinin. Strikingly, calretinin immunoreactivity was absent from spinal afferent neurons innervating the rat esophagus. In intraganglionic laminar endings and nodose ganglion cells calretinin was highly co-localized with calbindin but not with calcitonin gene-related peptide. On the other hand, calbindin was also found in spinal afferents to the esophagus where it was co-localized with calcitonin gene-related peptide. Vagal afferent neurons innervating the esophagus were never positive for NADPH diaphorase. Thus, calretinin appears to be a more specific marker for vagal afferent structures in the esophagus than calbindin, which is expressed by both vagal and spinal sensory neurons. Calretinin immunocytochemistry may be utilized as a valuable tool for investigations of subpopulations of vagal afferents in certain viscera.     

Variations in uterine innervation during the estrous cycle in rat

Uterine innervation undergoes profound remodeling during puberty, pregnancy, and after parturition. However, the extent to which uterine innervation may change during the estrous cycle is uncertain. The objective of the present study was to determine whether nerve fiber density of the uterine horn is altered during the estrous cycle and, if so, which subpopulations are affected. Immunostaining for the pan-neuronal marker protein gene product (PGP) 9.5 revealed fibers within the vascular zone, myometrium, and endometrium, with greater density in the ovarian and cervical regions than in the middle. In most structures, nerve density was reduced during estrus. This could not be accounted for by increased target volume, as the reduction in longitudinal muscle innervation persisted after correction for changes in target size. Immunostaining for vasoactive intestinal polypeptide-immunoreactive parasympathetic nerves revealed fibers associated predominantly with the vascular zone and circular muscle within the cervical region. No cyclical variation was detected. Calcitonin gene-related peptide-immunoreactive nerves were present within all structures, and density was highest at the ovarian end. These fibers also did not vary significantly through estrous. Dopamine beta-hydroxylase-immunoreactive sympathetic nerves innervated all structures, with greater density in the ovarian end. These fibers were reduced substantially during estrus, but the decline was also significant in proestrus, thus preceding that detected by using the pan-neuronal marker. We conclude that the estrous cycle in rat is accompanied by structural remodeling of sympathetic nerves by way of retraction or degeneration of terminal fibers during estrus. The structural loss of the terminal axon apparently is preceded by depletion of catecholamine-synthesizing enzyme.