Transforming Growth Factor α Evokes Aromatase Expression in Gastric Parietal Cells during Rat Postnatal Development.

Estrogen, well known as a female hormone, is synthesized primarily by ovarian aromatase. However, extra-glandular tissues also express aromatase and produce estrogen. It is noteworthy that aromatase in gastric parietal cells begins expression around 20 days after birth and continues secreting considerable amounts of estrogen into the portal vein throughout life, supplying it to the liver. Estrogen, which is secreted from the stomach, is speculated to play a monitoring role in blood triglyceride, and its importance is expected to increase. Nevertheless, the regulatory mechanisms of the aromatase expression remain unclear. This study investigated the influence of transforming growth factor α (TGFα) on gastric aromatase expression during postnatal development. The administration of TGFα (50 µg/kg BW) to male Wistar rats in the weaning period resulted in enhanced aromatase expression and increased phosphorylated ERK1+2 in the gastric mucosa. By contrast, administration of AG1478 (5 mg/kg BW), a protein tyrosine kinase inhibitor with high selectivity for the epidermal growth factor receptor and acting as an antagonist of TGFα, led to the suppression of aromatase expression. In fact, TGFα expression in the gastric fundic gland isthmus began around 20 days after birth in normal rats as did that of aromatase, which indicates that TGFα might induce the expression of aromatase in the parietal cells concomitantly.

Exploring heart dissection techniques for enhancing anatomical education: a pilot study to replicate transthoracic echocardiography.

PURPOSE: For novice learners, converting two-dimensional (2D) images of echocardiography to three-dimensional (3D) cardiac structures is deemed challenging. This study aimed to develop an accurate dissection method of the heart to reproduce the transthoracic echocardiographic views on cadavers and elucidate new educational methods in human anatomy dissection courses. METHODS: A total of 18 hearts were used in this study. After reflecting the anterior thoracic wall inferiorly, the hearts were excised from embalmed cadavers. Thereafter, three landmarks were set on the heart for each plane of the incision, and the hearts were incised to observe the three different echocardiographic views, which include the apical four-chamber view (A4C), parasternal long axis (PLAX) view, and parasternal short axis (PSAX) view at the papillary muscle level. If all structures for observation during routine echocardiography are clearly observed in each view, a successful incision is considered. All procedures and incisions were performed by the medical students. After a successful incision, hearts were returned to the original position in the pericardial sac for further observation. RESULTS: The success rates of incision for each view were 83.3% (5/6 success cases), 83.3% (5/6 success cases), and 66.7% (4/6 success cases) in the A4C view, PLAX view, and PSAX view at the papillary muscle level, respectively. CONCLUSION: This dissection method could probably be employed to reproduce transthoracic echocardiographic views on cadaveric hearts, which is beneficial for novice learners for a deeper understanding of the anatomy.

Anatomical analysis of ligaments surrounding calcaneocuboid joint; implications for role in foot stability.

PURPOSE: The calcaneocuboid joint is located in the lateral part of the foot and acts as a major stabilizer for the foot. Injuries to this joint often occur in association with ankle or foot injuries and are frequently overlooked, subsequently causing chronic pain or osteoarthritis. However, the relationship between ligaments surrounding the joint and joint instability remains unclear. Therefore, this study aimed to clarify the morphology and position of the ligaments surrounding the calcaneocuboid joint, and to reveal the relationship between the ligament structure. METHODS: The position and morphology of the bifurcate ligament (subdivided into calcaneonavicular and calcaneocuboid ligaments), dorsal calcaneocuboid ligament, lateral calcaneocuboid ligament, long plantar ligament, and short plantar ligament were measured (N = 11 feet in 6 Japanese cadavers). The circumference of the joint was quartered, while the ligament-uncovered area and the estimated cross-sectional area of each ligament were compared between the four sides. Furthermore, the estimated cross-sectional area of each ligament was calculated as an index for the ligament strength. RESULTS: The inferolateral side of the calcaneocuboid joint had the most uncovered area (54.63%) by the ligaments. In addition, the cross-sectional area of the ligaments on the lateral side was considerably smaller than that on the medial side. CONCLUSION: Our results suggest that ligament weakness on the inferolateral side may cause instability of the calcaneocuboid joint, especially after an inversion sprain injury, and may decrease the lateral longitudinal arch function, which results in chronic foot pain.

A bilateral third head of the gastrocnemius which is morphologically similar to the plantaris.

PURPOSE: An extra muscle was observed on both sides of the popliteal fossa in the cadaver of a 78-year-old Japanese male during dissection. The aim of this case report was to identify whether this variant is a double plantaris or a third head of the gastrocnemius according to its morphological characteristics and innervation. METHODS: The muscles were displayed by careful dissection and delineation of surrounding structures. The size of each of the muscle bellies and tendons of those extra muscles were measured manually by the vernier caliper. RESULTS: The origin of each extra muscle was lateral to the tibial nerve and superior to the plantaris, and each extra muscle which transitioned to a descending tendon parallel to the plantaris had a cone-shaped belly. However, the tendon of the extra muscles was fused into the investing fascia of the gastrocnemius with a tendon length of 4.5 cm on the left and 4.6 cm on the right. The extra muscles were innervated by the branch of the tibial nerve to the medial head of the gastrocnemius on both sides. CONCLUSION: Although they had an origin and shape similar to that of the plantaris, we identified the extra muscles in this case as a third head of the gastrocnemius, because of innervation to the plantaris arises directly from the tibial nerve. This case highlighted that the innervation is essential to understanding the myogenesis of extra muscles, especially in cases which are difficult to categorize based on the morphological features of the muscle.

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.

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.

A novel categorization of the muscular branches of the tibial nerve within the popliteal fossa.

BACKGROUND: The muscular branches of the tibial nerve within the popliteal fossa innervate the gastrocnemius, soleus, plantaris, and popliteus muscles. Various branching patterns have been described in textbooks; however, the underlying fundamental rules explaining the patterns remain unclear. Understanding the fundamental rule explaining the branching pattern of the innervating nerves is essential for understanding the ontogeny of skeletal muscles. Therefore, this study aimed at establishing a theory to explain the branching pattern of the muscular branches of the tibial nerve within the popliteal fossa. METHODS: The branching patterns of the muscular branches of the tibial nerve within the popliteal fossa were examined macroscopically in 62 lower limbs derived from 31 adult cadavers (22 males and 9 females, aged 49-95 years). RESULTS: The branch to the medial head of the gastrocnemius muscle invariably arose from the posteromedial side of the tibial nerve. The branches to the soleus muscle and lateral head of the gastrocnemius muscle had a common trunk in all the lower limbs and invariably arose from the posterolateral side. The branches to the plantaris and popliteus muscles arose anteriorly from the tibial nerve in this order (plantaris branch first, followed by the popliteus branch). These branches invariably arose more distally than the branch to both the heads of the gastrocnemius and soleus muscles. CONCLUSIONS: Based on these fundamental branching patterns, we suggest a novel branching categorization. The branches could be categorized into a posterior group and an anterior group, which has independent branches to the plantaris and popliteus muscles. This fundamental branching pattern and novel categorization contribute to the understanding of the ontogeny of the skeletal muscles around the flexor compartment of the leg.

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.

Tyrosine hydroxylase-immunoreactive fibers in the human vagus nerve.

Sympathetic catecholaminergic fibers in the vagus nerve were immunohistochemically examined in formalin-fixed human cadavers using an antibody against the noradrenalin-synthetic enzyme tyrosine hydroxylase (TH). TH-positive fibers were extensively distributed in the vagal nerve components, including the superior and inferior ganglia, the main trunk and the branches (superior and recurrent laryngeal, superior and inferior cardiac, and pulmonary branches). The inferior ganglion and its continuous cervical main trunk contained numerous TH-positive fibers with focal or diffuse distribution patterns in each nerve bundle. From these findings, we conclude that sympathetic fibers are consistently included in the human vagus nerve, a main source of parasympathetic preganglionic fibers to the cervical, thoracic and abdominal visceral organs.

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.

Functional role of lingual nerve in breastfeeding.

Functional role of lingual nerve in breastfeeding was investigated in rat pups during the suckling period. DiI, a postmortem neuronal tracer, was used to confirm the immature lingual nerve (LN) responsible for tongue sensation and resulted in successful fiber labeling anterogradely to the tongue, which showed different distribution patterns from fiber labeling derived from the hypoglossal nerve. Unilaterally LN-injured pups did not show suckling disturbance with absence of any shortening (P11 pups: 559+/-16s; 105% of the control value) in nipple attachment time and the survival rate remained high (P11: 100%). Bilaterally LN-injured pups showed suckling disturbance with marked shortening (P11 pups: 220+/-54 s; 42% of the control value) in nipple attachment time and a low survival rate (P1: 33%; P11: 41%). Bilaterally infraorbital nerve-injured or bilaterally bulbectomized pups did not show any nipple attachment at all and there were no survivors, confirming the crucial roles of upper lip sensation and olfaction in suckling. Based on these findings, we conclude that tongue sensation is very important, but not essential for suckling.

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.

Effect of maternal deprivation on milk intake in normal and bilaterally facial nerve-injured developing rats.

We examined effect of maternal deprivation (2, 4 and 6h) on milk intake in developing rats. Milk intake was obtained by body weight gain after 1h lactation. The amounts of milk intake significantly differed depending on the duration of the deprived periods at P7 and P14 with proportional increase by longer deprivation. Further, milk intake was measured in the bilaterally facial nerve-injured neonatal rats. The results show that milk intake is increased during development affected by maternal deprivation, and that milk intake of the facial nerve-injured group is decreased by 35% (2h), 7% (4h) and 7% (6h) at P7, and 25% (2h), 20% (4h) and 27% (6h) at P14 compared to that of the control group.

Description and first clinical application of AirWay Scope for tracheal intubation.

We have developed a novel airway apparatus, AirWay Scope, which we have used in the orotracheal intubation of 10 consecutive patients requiring general anesthesia before neurosurgical procedures. The characteristic shape of the introducer (INTLOCK), the tube guiding function and the sighting device of the AirWay Scope has possibility to facilitate tracheal intubation. Here, we report the first clinical application of this system and describe potential advantages of the apparatus.

The Relationship of the Subclavius Muscle with Relevance to Venous Cannulation below the Clavicle.

Introduction. The catheter "pinch-off syndrome" has been described to be secondary to crimping of the catheter between the clavicle and the first rib, as well as entrapment of the catheter at the site of penetration of the subclavius muscle. The lateral insertion technique has been recommended to prevent catheter pinch-off, but it is unknown if this technique can prevent entrapment by the subclavius muscle. We undertook this study to evaluate the anatomical relationship of the subclavius muscle and the subclavian vein. Methods. Twenty-eight adult cadavers were studied on both right and left sides. The adherence between the subclavian vein and subclavius muscle was subjectively assessed and the distance between the two structures was measured in mm. Results. The subclavius muscle and subclavian vein were tightly adherent in 72% of specimens, partly adherent in 14% with a mean distance of 4.5 mm and loosely connected in 14% with a mean distance of 6.1 mm. Conclusions. The anatomical relationship between the subclavius muscle and vein was very close in the majority of specimens, suggesting that the lateral insertion technique may not prevent penetration of the muscle, which may contribute to catheter pinch-off. The real-time ultrasound-guided technique may prevent penetration of the subclavius muscle.

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.

Neuregulin receptor ErbB2 localization at T-tubule in cardiac and skeletal muscle.

Previous studies have indicated that ErbB receptors for neuregulins play an important role in cardiac development and muscle spindle formation during embryogenesis; however, little is known about their functions in adulthood. Recent reports indicate that breast cancer therapy with humanized monoclonal ErbB2 antibody induces cardiomyopathy, suggesting that ErbB2 serves as a crucial signaling receptor, even in the adult heart. Here, we examine ErbB2 expression and localization in both cardiac and skeletal muscle of adult mice via immunoblot and immunohistochemistry. ErbB2 was detected as a band approximately 185 kD molecular mass in each cardiac and skeletal muscle extraction. Confocal images of double labeling showed that ErbB2 was colocalized with caveolin-3 in cardiac muscle and with dihydropyridine receptor in skeletal muscle, suggesting that ErbB2 was localized at the T-tubule. In addition, immunoelectron micrographs clearly demonstrated that ErbB2 was located at the T-tubule in both types of muscle. Taken together, the results of the present study suggest that neuregulin-ErbB2 signaling plays a role in the physiological function of cardiac and skeletal muscle, even in adulthood.

The minimum number of facial motor neurons essential for minimal whisker movement in neonatally nerve-transected young adult rats.

OBJECT: This study was performed to provide quantitative data on the number of surviving facial motor neurons that extend regenerated nerve fibers through the nerve-injured site and to reveal the relationship between facial function and the number of those motor neurons in which the facial nerve has been transected or resected in neonatal rats. METHODS: After transection of facial nerves in 1-day-old rat pups, facial function was estimated on postoperative Day 56 and a retrograde neuronal tracer was applied to the specific facial nerve branch responsible for the whisker movement. The mean number of the tracer-labeled neurons in the control rats was 2623+/-31 (mean+/-standard error of the mean) and that of the nerve-transected rats was 74+/-11 (range 0-221). Based on whisker movement, the nerve-transected rats were divided into two groups: clear spontaneous whisker movement and no whisker movement. The mean number of the tracer-labeled neurons in the nerve-transected rats with mobile whiskers was (106+/-12, range 44 [2% of the control value]-221 [8%]), whereas that in the nerve-transected rats with nonmobile whiskers was 24+/-6 (range 0-54 [2% of the control value]). The nerve-resected rats produced no labeled neurons. CONCLUSIONS: It was concluded that axotomized neonatal facial motor neurons extended regenerated nerve fibers through the nerve-transected site with the maximum value of 8% of the control value and that minimal whisker movement was preserved with a very small population of motor neurons (2%).

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.