Spontaneous regeneration after resection of various lengths of hypoglossal nerve in rats.

OBJECTIVES: The objective of this study was to investigate spontaneous neural regeneration and functional recovery after resection of various lengths of the hypoglossal (XII) nerve in adult rats. METHODS: Twelve weeks after XII nerve resection at lengths ranging from 0.0-15.8 mm, the tongue deviation angle of rats was measured to evaluate the severity of paralysis; thereafter, the XII neurons in the XII nucleus were labeled with Fluoro-Gold (FG), which was injected into the tongue to visualize regenerated XII neurons re-innervating the tongue muscles. RESULTS: In the XII nerve-resected rats, the regenerative rates, that is, the percentage of the total number of FG-positive neurons on the injured side relative to that on the uninjured side, were divided into two groups; the regenerative rates were more than 77% and less than 6%, respectively. Upon comparing the two groups, the boundary resection length was approximately 10.0 mm. Moreover, the former and latter groups demonstrated tongue deviation angles less than or greater than 15°, respectively. CONCLUSIONS: The critical nerve gap length for spontaneous neural regeneration was approximately 10.0 mm in XII nerve-resected adult rats, and nerve regeneration occurred in both morphological and functional aspects after resection at less than the critical length.

The calcaneofibular ligament groove at the inferior fibula, an ultrasonographic anatomical landmark.

PURPOSE: Calcaneofibular ligament (CFL) injuries are harder to diagnose than anterior talofibular ligament (ATFL) ones. This study aimed to clarify the fibular attachment of the CFL and verify the bony landmark for evaluating the CFL on ultrasonography. METHODS: Fifty-nine ankles were used in this anatomical study. To confirm the control function of the CFL, we performed passive movement manually using cadaveric ankles and observed the ankle positions where the CFLs were tense. Histological observation of CFL attachment of the fibula was performed using Masson's trichrome stain. The ATFL and CFL were removed, and the bone morphology of the CFL attachment and inferior fibular end was imaged using a stereomicroscope and a 3D scanner. Using ultrasonography, we evaluated the bone morphology of the fibular attachment of the CFL in short-axis images of 27 healthy adult ankles. RESULTS: The CFL was tensed according to ankle motions: supination, maximum dorsi flexion, maximum plantar flexion, and mild plantar flexion-external rotation. Below the CFL attachment of the fibula was a slight groove between the inferior tip and the obscure tubercle of the fibula. This groove was observed in 81.5% of cases using short-axis ultrasonography. CONCLUSION: The CFL was tensed in various ankle positions to control the movements of the talocrural and subtalar joints. There was a slight groove at the inferior end of the fibula where the CFL coursed downward. We called it the CFL groove and proposed that it could serve as a landmark for the short-axis image of ultrasonography.

Difference in the fibular attachment structure between the superior and inferior fascicles of the anterior talofibular ligament using ultrasonography and histological examinations.

PURPOSE: The anterior talofibular ligament (ATFL) is divided into superior (SB) and inferior bands (IB). Although the differences in length and width are known, the structure of the fibular attachment had not been elucidated. The present study aimed to clarify the differences in the fibular attachment structure between ATFL's SB and IB using cross-sectional images along the ligament. METHODS: An anatomical study using 15 formalin-fixed ankles was performed. The lateral ankle ligament complex was collected after a longitudinal image of SB/IB was visualized by ultrasonography. The specimens were decalcified and sectioned longitudinally at the center of SB/IB using a microtome. Histological evaluation of the enthesis structure at the fibular attachment of SB/IB was performed using hematoxylin-eosin and Masson's trichrome stains. RESULTS: A fibrillar pattern could not be observed in the longitudinal image at the IB level by ultrasonography. The lengths of ATFL's SB and IB were 20.6 ± 1.6 and 15.3 ± 1.3 mm, respectively, with thicknesses of 1.8 ± 0.4 and 1.0 ± 0.4 mm, respectively. The ATFL's IB was significantly shorter and thinner than the ATFL's SB. The fibular attachment of ATFL's SB had distinct enthesis structure, whereas in the attachment structure of the ATFL's IB, there were several variations including a type with a narrower enthesis structure than the ATFL's SB and a type that merged with or wrapped around the calcaneofibular ligament. CONCLUSION: The fibular attachment structure between ATFL's SB and IB differs. Our results could be useful information when performing ultrasonography and MRI diagnosis.

Relationship between inferior fascicle of anterior talofibular ligament and articular capsule in lateral ankle ligament complex.

PURPOSE: The lateral ankle ligament complex (LALC) is composed of anterior talofibular (ATFL), calcaneofibular (CFL), and posterior talofibular (PTFL) ligaments, all of which have a connection/continuous fiber. However, the structural link between the LALC and the articular capsule remains unknown. The goal of our study was to determine the connection between ATFL's inferior fascicle and the articular capsule. METHODS: In this study, we utilized 84 formalin-fixed ankles to elucidate the structure of LALC. Between ATFL and CFL, the bundle number of ATFL and arciform fiber was investigated. The specimens were decalcified and sectioned coronally using a freezing microtome, in the case of double bundles of ATFL, to study the connection between the inferior fascicle of ATFL and the articular capsule. RESULTS: ATFL had a single (25%), double (74%), and triple (1%) bundle number, respectively. The arciform fiber connecting the ATFL and the CFL was found in the superficial layer of all ankles (100%). There were two types of relationships between the inferior fascicle of ATFL and the articular capsule: 36 ankles (58%) were extracapsular, and 26 of 62 ankles (42%) were integrated with the inferior-lateral articular capsule. There are two kinds of relationships between the inferior fascicle of the ATFL and the articular capsule: extracapsular and integrated-capsular. CONCLUSIONS: The inferior fascicle of ATFL has a variant and integrated-capsular type is reinforced inferior-lateral articular capsule and enters the joint to form continuous fibers with PTFL, making LALC. These anatomical findings are helpful in ultrasonography diagnosis and arthroscopic ankle surgery.

Continuous and Connective Fibers of the Lateral Ankle Ligament Complex.

The lateral ankle ligament complex (LALC) is an intricate structure; therefore precise anatomic knowledge is required by the surgeon. However, the structural relationship of the LALC remains unclear. Here, the features of the posterior talofibular ligament (PTFL) and the relationship to the LALC at the distal fibula were clarified in a cadaver study. The lengths of most of the anterior and posterior parts, and the widths of the anterior-posterior and superior-inferior parts, were measured with a digital caliper. In addition, the relationship between the anterior talofibular ligament (ATFL) and PTFL inside of the capsule is described. The small fiber bundles of the PTFL were manually divided, and the footprint of each bundle at the fibula and talus was clarified. The relationship between the ATFL and CFL, outside of the capsule, was examined on axial slices at the inferior fibula. The lengths of the most anterior and most posterior parts of the PTFL were 9.8 ± 1.7 and 29.4 ± 1.9 mm, respectively. The widths of the anterior-posterior and superior-inferior parts were 10.0 ± 0.9 and 5.8 ± 1.1 mm, respectively. Approximately 83% of the fibers between the ATFL and PTFL were continuous. The anterior-inferior fibers of the PTFL were continuous with the inferior fibers of the ATFL inside of the capsule. The ATFL and CFL converged with connective tissue from outside of the capsule at the distal fibula. The results of this study should prove useful to further clarify the relationships of the LALC both inside and outside of the capsule at the distal fibula.

Hypoglossal nerve injury with long nerve resection leading to slow motoneuron death.

Crush injury to peripheral nerves in adult animals is considered not to trigger retrograde neuronal cell death; however, several studies reported neuronal cell death following severe injuries including nerve transection, resection, or avulsion. However, the rate of neuronal cell death varied among studies. In this study, we evaluated the outcomes of very severe nerve injury by long nerve resection in adult rats. Right hypoglossal (XII) nerve was exposed, and a 9-mm section was resected. At 4, 8, and 12 weeks after the resection, the number of XII neurons were counted in from the rostral to caudal sections. The number of XII neurons in the injured right side was reduced after the XII nerve resection compared with the uninjured left side. The mean rates of surviving neurons at 4, 8, and 12 weeks after the nerve resection were 83.5 %, 73.9 %, and 61.1 %, respectively, which were significantly lower than those of the control. The number of XII neurons after extensive XII nerve resection declined gradually over a relatively long time period, revealing that extensive nerve resection led to slow cell death of the injured neurons.

The interrelationship between anterior cruciate ligament tibial footprint and anterolateral meniscal root insertions: Quantitative, morphological and positional analyses using three-dimensional computed tomography images.

BACKGROUND: The purpose of this study was to evaluate quantitative, morphological and positional differences between the anterior cruciate ligament (ACL) tibial footprint and anterolateral meniscal root (ALMR) insertion and investigate an intraoperative landmark to estimate their boundaries. METHODS: Thirty-three fixed human cadaveric knees were evaluated. After resecting the components, the anterior fiber (AF) and posterior fiber (PF) of ALMR, the tibial center of ACL bundles (anteromedial (AM) and posterolateral (PL) bundles) and ACL were marked. Insertion morphology was classified into three categories, and the distance and relative positional relationship between AF/PF insertions and the center of each attachment were measured on three-dimensional computed tomography images. RESULTS: There was no significant difference between the AF of AM and ACL (P = 0.16), but both were significantly shorter than the AF of PL (both P < 0.001). There was no significant difference between the PF of ACL and PL (P = 0.99), which were significantly shorter than PF of AM (both P < 0.001). Morphology of the ACL tibial insertion was classified as follows: triangular, 15 knees (45.5%); oval, 18 knees (54.5%); none, C-shape. Quantitative and positional analyses showed that the AF insertion was significantly closer to AM and ACL centers in the oval type than in the triangular type. Excluding two cases, the AF/PF insertion was located laterally to the ML center of the medial and lateral intercondylar tubercles. CONCLUSION: Proximity of ACL tibial footprint and ALMR varies by their footprint morphology. The medial and lateral intercondylar tubercles were useful landmarks for ALMR injury prevention.

Independent Attachment of Lateral Ankle Ligaments: Anterior Talofibular and Calcaneofibular Ligaments - A Cadaveric Study.

Anatomic knowledge of lateral ligaments around the lateral malleolus is important for repair or reconstruction of ankle instability. The detailed structure of the connective fibers between the anterior talofibular ligament (ATFL) and the calcaneofibular ligament (CFL) is unknown. To clarify the anatomic structure of ATFL and CFL and the connective fiber between the 2 ligaments, the lateral ligament was dissected in 60 ankles of formalin-fixed cadavers, and the distance was measured between bony landmarks and fibular attachment of ATFL and CFL using a digital caliper. All ankles had connective fibers between ATFL and CFL. The structure of connective fibers consisted of a thin fiber above the surface layer of ATFL and CFL; it comprised thin fibrils of the surface layer covering the lower part of ATFL and the front part of CFL. Both ATFL and CFL were independent fibers, and both attachments of the fibula were isolated. Single bands of ATFL were noted in 14 of 60 (23.3%) ankles, double bands that divided the superior and inferior bands were observed in 42 of 60 (70.0%) ankles, and multiple bands were observed in 4 of 60 (6.7%) ankles. A cord-like and a flat and fanning type of CFL was noted in 22 (36.7%) and 38 (63.3%) of the 60 ankles, respectively. Distances between ATFL/CFL and articular and inferior tips of the fibula were 4.3 ± 1.1 mm/7.6 ± 1.6 mm and 14.3 ± 1.9 mm/7.4 ± 1.7 mm, respectively (mean ± standard deviation). The results of this study suggest that knowledge of more anatomic structures of ATFL, CFL, and connective fiber will be beneficial for surgeons in the repair or reconstruction of the lateral ligament of the ankle.

Effects of various lengths of hypoglossal nerve resection on motoneuron survival.

We employed stereological analyses for whole quantification of hypoglossal (XII) motoneurons in adult rats that received varying degrees of resection of the XII nerve. Various lengths of nerve gaps (0.0-13.3 mm) were made at the main trunk of the unilateral XII nerve, and the total number of XII neurons on the injured and uninjured sides was counted 12 weeks after nerve resection. The stereologically estimated total number of XII neurons decreased after various lengths of nerve resection, and survival rates ranged from 34.4% to 87.1%. Statistically significant negative correlations were observed between increasing length of the resected nerve and decreasing XII neuron survival. It was concluded that the total number of XII neurons decreased after nerve resection and that survival rates of XII neurons were related to distances between resected nerve stamps.

Effects of repeated nerve injuries at different time intervals on functional recovery and nerve innervation.

Effects of repeated nerve injuries on functional recovery and nerve innervation were examined in rodents. Crush injuries of the sciatic nerve were inflicted on adult rats and repeated twice or thrice at different time intervals of 1, 2, 3, and 4 weeks. Motor function was assessed by the static sciatic index at 1, 7, 14, 21, 28, 35, 42, 49, and 56 days after the final crush. The rates of nerve innervation of the tibialis anterior muscle, a main muscle innervated by the common peroneal nerve, were evaluated by the quantification of ßIII-tubulin-positive nerve terminals and α-bungarotoxin-positive acetylcholine receptors 21 and 56 days after the final crush of triple nerve injuries at 1-, 2-, 3-, and 4-week intervals. Compared with single nerve crush injury, delayed recovery of motor function was observed in repeated crush injuries. In addition, recoveries in the triple crush groups were slower than those in the double crush groups. The rates of reinnervation were lower in the triple crush groups than in the single crush groups, both at 21 days (single: 59.7%; triple: 54.1%-56.1%) and 56 days (single: 88.8%; triple: 72.5%-83.0%) after the final crush, except in the groups with 1-week (triple: 73.8%) and 2-week (triple: 70.5%) intervals at 21 days after the final crush. We concluded that the recovery of motor function was delayed according to the number of repetitions of crush injuries, and that the rates of nerve innervation were still low in the triple crush groups 8 weeks after the final crush.

Acetyl-l-carnitine enhances myelination of regenerated fibers of the lateral olfactory tract.

It is well known that acetyl-l-carnitine (ALC) has various neuroprotective effects against neurodegenerative diseases. In addition, it has been reported that ALC facilitates myelination of regenerated axons after peripheral nerve injuries. We previously reported that spontaneous regeneration of the lateral olfactory tract (LOT), the main fiber tract of the central olfactory system, consistently occurred in newborn rats and a majority of these regenerated fibers were unmyelinated in neonatally LOT-transected young adult rats. To investigate the effects of ALC treatment on myelination in LOT, neonatal rats were treated with ALC after LOT transection. Immunohistochemistry for myelin basic protein showed more positive areas in ALC-treated rats than in control rats. Moreover, the number of myelinated axons of regenerated fibers was assessed using electron microscopy and was found to be statistically higher in ALC-treated rats compared to control rats. The study revealed that ALC accelerates myelination of regenerated fibers in neonatally LOT-injured young adult rats.

Stereological assessment of the total number of hypoglossal neurons after repeated crush injuries to the hypoglossal nerve in adult rats.

OBJECTIVE: Retrograde neuronal cell death does not occur in mature motoneurons following the axonal injury of peripheral nerves. However, a previous study suggested that retrograde neuronal cell death does occur in adult rats after the creation of double lesions on the hypoglossal (XII) nerve based on a substantial decrease in the number of XII neurons. Using stereological methods, we examined neuronal apoptosis in XII neurons and the total number of XII neurons following repeated crush injuries to the XII nerve. METHODS: The right XII nerve of adult rats was crushed three times at one-week intervals with a brain aneurysm clip. At 4 weeks after the final crush, the total numbers of XII neurons on the injured right and uninjured left sides were estimated stereologically. RESULTS: After repeated crush injuries, no apoptosis was evident in XII neurons as indicated by immunostaining for cleaved caspase-3. Moreover, immunohistochemistry for the vesicular acetylcholine transporter revealed axonal elongation in the tongue 4 weeks after repeated crush injuries. At 4 weeks, the total numbers of XII neurons were 7800 ± 290 on the injured right side and 8000 ± 230 on the uninjured left side, and no significant difference was evident between the injured and uninjured sides. CONCLUSION: Neuronal cell death does not occur in XII neurons and the total number of XII neurons does not decrease after repeated crush injuries of the XII nerve in adult rats.

Morphological analysis of regenerated bulbar fibers in relation to neonatal olfaction.

It was revealed that regeneration of the lateral olfactory tract (LOT) occurred in developing rats and the regenerated olfactory system was functional 4 weeks after transection. The aim of this study was to determine the earliest onset of functional recovery in LOT-injured rats and to quantify regenerated nerve components with functional correlation. Neonatal rats on postnatal day (P) 2 were subjected to unilateral transection of the left LOT and underwent unilateral removal of the right olfactory bulb on P11. Functional recovery of the tract injury was assessed by the suckling capability, which can be achieved by olfaction. Suckling capability was observed on P12 in most neonatally LOT-transected pups. Rat pups were subjected to unilateral transection of the left LOT on P2, and received injections of biotinylated dextran amine (BDA) into the bilateral olfactory bulb on P5 to quantify normal and regenerated nerve components in the olfactory cortices at the level of the olfactory tubercle. BDA(+) areas and density indices of the olfactory cortices in the neonatally LOT-transected P12 pups were 11.05×105µm2 and 0.35 on the normal right side and 4.34×105µm2 and 0.21 on the transected left side. We concluded that functional recovery of the LOT-transected neonatal rats occurred as early as 10days after tract transection and that areas and densities of regenerated nerve components essential for functional recovery were approximately 40% and 60% of the age-matched normal values in the olfactory cortices at the level of the olfactory tubercle.

Effect of graded nerve pressure injuries on motor function.

OBJECT: The purpose of this study was to determine the minimum amount of nerve fibers required to maintain normal motor function after nerve injury in rats. METHODS: The authors first confirmed that a common peroneal nerve injury caused more aggravating effects on lower limb motor function than tibial nerve injury, as assessed by the static sciatic index (SSI). Thereafter, rats were subjected to varying degrees of crush injury to the common peroneal nerve. At 48 hours after the injury, motor function was assessed using the SSI and slope-walking ability (with slope angles of 30° and 45°). The tibialis anterior muscle, a main muscle innervated by the common peroneal nerve, was removed. Muscle sections were co-labeled with neuronal class III ß-tubulin polyclonal antibody to identify the presence of axons and Alexa Fluor 488-conjugated α-bungarotoxin to identify the presence of motor endplates. RESULTS: The evaluation of neuromuscular innervation showed a correlation between SSI scores and ratios of residual axons (rs = 0.68, p < 0.01), and there was a statistically significant difference between slope-walking ability and ratios of residual axons (p < 0.01). Moreover, the ratios of residual axons in the nerve-crushed rats with normal motor function (SSI above -20) ranged from 36.5% to 88.7%, and those ratios in the success group with slope-walking angles of 30° and 45° ranged from 14.7% to 88.7% and from 39.8% to 88.7%, respectively. CONCLUSIONS: In this study of rodents, less than half of the motor axons were sufficient to maintain normal motor function of the lower limb.

Stereological estimation of olfactory receptor neurons in rats.

The total number of olfactory receptor neurons (ORNs) in the rat main olfactory epithelium (MOE) was estimated using stereological sampling. Skulls and noses of newborn (postnatal day 0), young adult (8 weeks), and adult (6 months) rats were decalcified, embedded in paraffin, cut into 10-µm thick sections serially at 100-µm intervals, and processed for immunohistochemistry for olfactory marker protein (OMP), a specific marker of mature ORNs with fiber connections to the olfactory bulb. The number of OMP (+) receptor neurons was measured using an optical fractionator with stereological software (Stereoinvestigator). The total number of OMP (+) receptor neurons in the unilateral MOE was 0.47 × 10(6) in newborns and 21 × 10(6) in young adults and adults. Our previous study showed that the corresponding unilateral total number of neurons was 5.2 × 10(6) in young adult mice. Accordingly, we concluded that rats had 4 times more OMP (+) receptor neurons than mice at the adult stage and that the number of these neurons increased approximately 45 times between birth and maturity.

Two specific tongue regions receive bilateral hypoglossal innervation: a study in neonatal rat pups.

OBJECTIVE: The purpose of this study was to investigate the functional role of bilateral hypoglossal (XII) nerve innervation of the tongue. MATERIALS AND METHODS: The possibility of bilateral innervation of XII nerve in the tongue was examined using postmortem fibre tracing in normal neonatal rats. In addition, immunohistochemical testing for synaptophysin or vesicular acetylcholine transporter (VAChT) was carried out in unilaterally XII nerve-resected neonatal rats. RESULTS: Postmortem fibre tracing revealed constant distribution of the nerve fibres extending across the midline and existence of bilateral innervated area in the rostroventral and caudodorsal regions of the tongue. Synaptophysin-positive and VAChT-positive nerve terminals were also distributed continuously from the XII nerve-intact to the nerve-resected side across the midline of the tongue. The contralaterally projecting VAChT-positive nerve terminals were more numerous in suckling P2 rats (6.6 ± 0.5/section) than those in non-suckling P2 rats (4.9 ± 0.3/section) 24 h after nerve resection. Furthermore, the contralaterally projecting VAChT-positive nerve terminals were more numerous in P7 rats with nerve resection on P1 (6.3 ± 0.2/section) than those in P7 rats with nerve resection on P6 (3.1 ± 0.8/section). CONCLUSION: We concluded that neonatal rats have two specific tongue regions receiving bilateral XII innervation, which allowed suckling in unilaterally XII nerve-resected neonatal rats.

Correlation between motor function and axonal morphology in neonatally sciatic nerve-injured rats.

The present study was conducted to investigate the correlation between motor function and axonal morphology in neonatally sciatic nerve-injured rats. The left sciatic nerve of newborn rats was transected or crushed, and functionality of the sciatic nerve was assessed by the static sciatic index after 8 weeks. After functional assessment, the common peroneal nerves in the control, nerve-transected, and nerve-crushed rats were removed and prepared for morphometric examinations. The cross-sectional area of the nerve, total number of myelinated axons, and size of each myelinated axon were analyzed for each group. The control rats showed normal motor function, whereas the nerve-transected rats showed severe motor dysfunction. The cross-sectional area of the nerve and total number of myelinated axons were reduced after nerve transection. Moreover, the percentage per size class of myelinated axons was almost uniform in the control rats, while the distribution was shifted to the left in the nerve-transected rats. Furthermore, no large myelinated axons were observed in the nerve-transected rats. The nerve-crushed rats showed various gait functions with various distribution patterns of axonal size, and the rats were divided into two groups with and without uninjured residual large axons. The results showed that the importance of regenerated medium-sized axons in cases without large axons and of residual large axons in cases with large axons in motor function. It was revealed that motor function was related closely to axonal size in neonatally nerve-injured rats.

Bony Landmarks of the Anterior Cruciate Ligament Tibial Footprint: A Detailed Analysis Comparing 3-Dimensional Computed Tomography Images to Visual and Histological Evaluations.

BACKGROUND: Although the importance of tibial tunnel position for achieving stability after anterior cruciate ligament (ACL) reconstruction was recently recognized, there are fewer detailed reports of the anatomy of the tibial topographic footprint compared with the femoral side. HYPOTHESIS: The ACL tibial footprint has a relationship to bony prominences and surrounding bony landmarks. STUDY DESIGN: Descriptive laboratory study. METHODS: This study consisted of 2 anatomic procedures for the identification of bony prominences that correspond to the ACL tibial footprint and 3 surrounding landmarks: the anterior ridge, lateral groove, and intertubercular fossa. In the first procedure, after computed tomography (CT) was performed on 12 paired, embalmed cadaveric knees, 12 knees were visually observed, while their contralateral knees were histologically observed. Comparisons were made between macroscopic and microscopic findings and 3-dimensional (3D) CT images of these bony landmarks. In the second procedure, the shape of the bony prominence and incidence of their bony landmarks were evaluated from the preoperative CT data of 60 knee joints. RESULTS: In the first procedure, we were able to confirm a bony prominence and all 3 surrounding landmarks by CT in all cases. Visual evaluation confirmed a small bony eminence at the anterior boundary of the ACL. The lateral groove was not confirmed macroscopically. The ACL was not attached to the lateral intercondylar tubercle, ACL tibial ridge, and intertubercular space at the posterior boundary. Histological evaluation confirmed that the anterior ridge and lateral groove were positioned at the anterior and lateral boundaries, respectively. There was no ligament tissue on the intercondylar space corresponding to the intercondylar fossa. In the second investigation, the bony prominence showed 2 morphological patterns: an oval type (58.3%) and a triangular type (41.6%). The 3 bony landmarks, including the anterior ridge, lateral groove, and intertubercular fossa, existed in 96.6%, 100.0%, and 96.6% of the cases, respectively. CONCLUSION: There is a bony prominence corresponding to the ACL footprint and bony landmarks on the anterior, posterior, and lateral boundaries. CLINICAL RELEVANCE: The study results may help create an accurate and reproducible tunnel, which is essential for successful ACL reconstruction surgery.

Quantitative analysis of survival of hypoglossal neurons in neonatally nerve-injured rats: Correlation with milk intake.

INTRODUCTION: Tongue movement innervated by the hypoglossal (XII) nerve is essential for the survival of neonatal rats. The pups with bilateral XII nerve resection failed to suckle milk and did not survive, and the pups with unilateral XII nerve resection showed disturbed suckling capability and lower survival rates. The present study was performed to investigate the relation between neuronal population and milk intake of developing rats that had received various degrees of crush injuries to the unilateral XII nerve during the neonatal period. METHODS: The right XII nerve of postnatal day 1 (P1) pups was crushed and milk intake was estimated at 3 days and 6 days after the nerve injury. As nerve injury at the neonatal stage results in death of axotomized neurons, varying degrees of crushing was estimated by the number of survived motor neurons. RESULTS: In nerve-crushed rats, the populations of XII motor neurons and amounts of milk intake were reduced in a varied manner. Statistically significant positive correlations were observed between increasing XII neuron survival and increasing milk intake at 3 (r=0.62) and 6 (r=0.71) days after the nerve injury. CONCLUSION: The results indicate that there is a strong relationship between the number of XII motor neurons and the amount of milk intake in neonatally XII nerve-injured rats.

Regenerative capacity of bulbar projection neurons during development: a quantitative neuronal analysis with functional correlation.

Critical periods and degrees of regeneration in injured olfactory bulbar projection neurons (mitral cells) were examined in adult rats whose lateral olfactory tracts (LOTs) were transected at different postnatal (P) days. After the LOTs were transected at P7, P10, and P14, a retrograde fluorescent tracer, Fluoro-Gold (FG), was injected into the posterior olfactory cortex (the olfactory tubercle and the piriform cortex), a target brain region of mitral cells, 5 weeks after the transection. FG (+) mitral cells were observed in P7 LOT-transected bulbs and some of P10 LOT-transected bulbs but not in P14 LOT-transected bulbs. Neuron numbers of regenerated FG (+) mitral cells in P2 LOT-transected adult rats decreased to approximately 70% of the normal values (actually counted number: 804±46; stereologically estimated number: 49 700±4300), and 100% of these rats were demonstrated to exhibit olfactory discriminative ability in our previous study. Meanwhile, the numbers in P7 LOT-transected adult rats further decreased to approximately 40% of the normal values, and 78% of these rats showed olfactory discriminative ability. We conclude that the critical periods of spontaneous regeneration of the LOT are between P0 and P10 and that the proportions of regenerated mitral cells decreased as rats became older.