Location, size and age at onset of metamorphosis in the Japanese eel Anguilla japonica.

This study clarifies the location, size and age at the onset of metamorphosis in Japanese eels Anguilla japonica through oceanic surveys, rearing experiments and analyses of the morphology and otoliths of leptocephali and glass eels. Twenty-eight metamorphosing leptocephali were collected in the mesoscale eddy region to the east of Taiwan during research expeditions in 2004. Rearing experiments showed that the total length (LT ) of leptocephali decreased by an average of 12·5% during metamorphosis and 13·9% during the 2-12 h after death. Thus, the mean back-calculated LT at the onset of metamorphosis for 630 glass eels from Taiwan and Japan was estimated at 67·8 ± 2·7 mm (mean ± S.D.). The estimated mean ante-mortem size of the fully grown pre-metamorphic leptocephali collected in 2004 was 64·6 ± 3·4 mm, which was consistent with the LT estimate for glass eels. Otolith analysis showed that the mean age at the onset of metamorphosis was 137 ± 15 days and indicated that Japanese eels may have a recruitment route through the mesoscale eddies to the east of Taiwan in addition to the direct transfer route from the North Equatorial Current to the Kuroshio Current.

Stereological quantification of olfactory receptor neurons in mice.

The total number of olfactory receptor neurons (ORNs) in the mouse main olfactory epithelium (MOE) was estimated using stereological sampling. Noses and skulls of male and female 8-week-old C57BL/6J mice were de-calcified, embedded in paraffin, cut into 10-µm-thick sections serially at 100-µm intervals, and processed for immunohistochemistry for the olfactory marker protein (OMP), a specific marker for ORNs. The number of OMP (+) receptor neurons was measured using an optical fractionator with the Stereo-Investigator software. The mean values of the total number of OMP (+) receptor neurons in the unilateral MOE were 5,140,000±380,000 in males and 5,210,000±380,000 in females, with no significant differences between the sexes. We concluded that the total number of ORNs in the unilateral MOE is approximately 5×10(6) in mice.

Head-shape polymorphism in Japanese eels Anguilla japonica in relation to differences of somatic growth in freshwater and brackish habitats.

The age, total length (L(T)), head shape and skull shape were investigated for 379 Japanese eels Anguilla japonica sampled in freshwater and brackish areas of the Kojima Bay-Asahi River system, Okayama, Japan, to learn about the differentiation process of head-shape polymorphism. The relative mouth width (ratio of mouth width to L(T)) of A. japonica > 400 mm L(T) collected in fresh water was significantly greater than that of fish collected in brackish water. Growth rates of mouth width and the distance from the snout to the midpoint of the eyes (the ratio of width and distance to age, respectively) were not significantly different between freshwater and brackish-water samples, whereas the somatic growth rate (the ratio of L(T) to age) of freshwater samples was significantly lower than that of brackish-water eel samples. These results suggest that the factors affecting head and somatic growth of A. japonica are not identical. According to these results and feeding patterns in each habitat reported by another study, it is suggested that somatic growth appears to play a significant role in the differentiation process of the head-shape polymorphism in A. japonica, with the slow-growing fish in fresh water becoming broad-headed and the fast-growing fish in brackish water becoming narrow-headed.

Identification of a 3.7-Mb region for a marbling QTL on bovine chromosome 4 by identical-by-descent and association analysis.

QTL mapping for growth and carcass traits was performed using a paternal half-sib family composed of 325 Japanese Black cattle offspring. Nine QTL were detected at the 1% chromosome-wise significance level at a false discovery rate of less than 0.1. These included two QTL for marbling on BTA 4 and 18, two QTL for carcass weight on BTA 14 and 24, two QTL for longissimus muscle area on BTA 1 and 4, two QTL for subcutaneous fat thickness on BTA 1 and 15 and one QTL for rib thickness on BTA 6. Although the marbling QTL on BTA 4 has been replicated with significant linkages in two Japanese Black cattle sires, the three Q (more marbling) haplotypes, each inherited maternally, were apparently different. To compare the three Q haplotypes in more detail, high-density microsatellite markers for the overlapping regions were developed within the 95% CIs (65 markers in 44-78 cM). A detailed haplotype comparison indicated that a small region (<3.7 Mb) around 46 cM was shared between the Qs of the two sires, whose dams were related. An association of this region with marbling was shown by a regression analysis using the local population, in which the two sires were produced and this was confirmed by an association study using a population collected throughout Japan. These results strongly suggest that the marbling QTL on BTA 4 is located in the 3.7-Mb region at around 46 cM.

Depth distribution and biological characteristics of the European eel Anguilla anguilla in Lough Ennell, Ireland.

Using a longline survey, a total of 196 European eels Anguilla anguilla were collected at different depths in Lough Ennell (maximum depth 30 m), central Ireland. The catch per unit of effort of A. anguilla that were caught from 1 to 25 m depths was lowest at 0.5-5.0 m and greatest at the deepest depth range (22.5-25.0 m). Sub-samples of A. anguilla from depths of <15 m showed little or no difference in size, sex ratio, age, growth rate, condition factor, length-mass relationship, gonado-somatic index, fin index or eye index with fish from depths of >15 m. All fish examined were female yellow-phase A. anguilla that had ages from 7 to 20 years (mean +/-s.d. = 10.3 +/- 2.9 years), with growth rates from 24.0-60.8 mm year(-1) (mean +/-s.d. = 40.7 +/- 8.5 mm year(-1)). Variations in the growth rates were greater in the shallow group than that of the deep group. This study suggested that deeper regions are important feeding habitats for A. anguilla and that fish in this lake were growing moderately fast compared to similar habitats and areas in the species' range.

Linkage mapping of the locus responsible for forelimb-girdle muscular anomaly of Japanese black cattle on bovine chromosome 26.

Forelimb-girdle muscular anomaly is an autosomal recessive disorder of Japanese black cattle characterized by tremor, astasia and abnormal shape of the shoulders. Pathological examination of affected animals reveals hypoplasia of forelimb-girdle muscles with reduced diameter of muscle fibres. To identify the gene responsible for this disorder, we performed linkage mapping of the disorder locus using an inbred pedigree including a great-grand sire, a grand sire, a sire and 26 affected calves obtained from a herd of Japanese black cattle. Two hundred and fifty-eight microsatellite markers distributed across the genome were genotyped across the pedigree. Four markers on the middle region of bovine chromosome 26 showed significant linkage with the disorder locus. Haplotype analysis using additional markers in this region refined the critical region of the disorder locus to a 3.5-Mb interval on BTA26 between BM4505 and MOK2602. Comparative mapping data revealed several potential candidate genes for the disorder, including NRAP, PDZD8 and HSPA12A, which are associated with muscular function.

Differential effects of hypoglossal and facial nerve injuries on survival and growth of rats at different developmental stages.

The hypoglossal (XII) nerve is made up of functionally different nerve branches: the medial branch related to protrusion of the tongue and the lateral branch related to its retraction. The present study was performed to determine the effects of facial (VII) and XII nerve injuries on the survival and growth of rats in which the unilateral or bilateral VII and XII nerve components (main trunk, XII-trunk; medial branch, XII-med; lateral branch, XII-lat) had been resected at different developmental stages. In the suckling period, unilateral as well as bilateral injuries in the XII-trunk or XII-med nerve produced disturbed milk intake, lower survival rates and growth retardation in the nerve-injured rats. In the transition and mastication periods, only bilateral injury in the XII-trunk or XII-med nerve produced disturbed food intake followed by lower survival rates and growth retardation in those animals. The unilateral XII-lat nerve injury did not have significant effects on milk and food intake, whereas the bilateral injury caused disturbance in milk intake especially at the early neonatal stage. The unilateral VII nerve injury at the early neonatal stage caused deteriorating effects on food intake resulting in lower survival rate and severe growth retardation in the nerve-injured rats. The results indicate that the survival and growth of XII and VII nerve-resected rats differ considerably depending on the nerves injured and the developmental ages of the animals at the time of nerve insult.

Genome-wide linkage disequilibrium in two Japanese beef cattle breeds.

There is little knowledge about the degree of linkage disequilibrium (LD) in beef cattle. This study aims to perform a genome-wide search for LD in Japanese Black and Japanese Brown beef cattle and to compare the level of LD between these two breeds. Parameter D' (the LD coefficient) was used as a measure of LD, and LD was tested for significance of allelic associations between syntenic and between non-syntenic marker pairs. Effects of breed, chromosome, genetic map distance and their interactions with D' were tested based on least squares analyses. Both breeds showed high levels of LD, which ranged over several tens of cM and declined as the marker distance increased for syntenic marker pairs. A rapid decline of the D' value was observed between markers that were spaced 5 and 20 cM apart. LD was significant in most cases for marker pairs <40 cM apart but was not significant between non-syntenic loci. The pattern of LD found in these two breeds was similar to that previously published for dairy cattle. The D' value between breeds was not significantly different (P > 0.05), but the interaction between breed and chromosome was highly significant (P < 0.001). Genetic selection seems to have caused the heterogeneity of the D' values among chromosomes within breed. These results indicate that LD mapping is a useful tool for fine-mapping quantitative trait loci of economically important traits in Japanese beef cattle.

Effects of hypoglossal and facial nerve injuries on milk-suckling.

Functional roles of the perioral anatomical structures involved in breastfeeding were examined in newborn rat pups in which the hypoglossal (XII) and facial (VII) nerves had been resected at the neonatal stage. The XII nerve controls tongue movement and is comprised of two functionally distinct branches: the medial branch related to protrusion of the tongue and the lateral branch related to its retraction. Newborn rat pups with bilateral resection of either of the XII nerve components (main trunk: XII-trunk; medial branch: XII-med; lateral branch: XII-lat) failed to suckle milk and did not survive. Unilateral XII nerve-resected neonates showed different milk-suckling capabilities, which thus resulted in differences in survival rate (XII-trunk: 38%; XII-med: 24%; XII-lat: 92%) and postnatal growth during the postnatal 3 weeks until P21. Unilateral and bilateral resections of the VII nerve innervating the buccolabial musculature produced lowered suckling capabilities and retarded postnatal growth, although all pups showed 100% survival. The results indicate a crucial role of the tongue, especially of protruding muscular elements innervated by the XII-med nerve, in breastfeeding. The results also indicate differential effects of the VII and XII nerve components on suckling capability, survival, and postnatal growth of newborn rat pups.

Effects of bilateral resection of facial nerves on suckling in developing rats.

The purpose of the present study is to investigate functional role of the facial nerve on suckling in developing rats. The bilateral resection of facial nerves on postnatal day 1 (P1) resulted in cell loss of facial motoneurons and complete facial paralysis without any whisker movement or nictitating reflex at the end of the postnatal 3 weeks. Although the body weight of the nerve-resected rats increased gradually for the postnatal 3 weeks, they weighed less than the control rats without nerve resection. The nerve-resected rats contained less milk (0.25 +/- 0.02 g) than the control rats (0.35 +/- 0.02 g) in the stomach on P17. On P21, the body weight of the nerve-resected rats (25.33 +/- 0.32 g) was decreased by 28% compared to that of the control rats (35.08 +/- 0.57 g). Although their growth was substantially more retarded than that of the control animals, most (92%) of the nerve-resected pups could survive without facial nerve innervation. The orofacial musculature innervated by the facial nerve plays an important role in breastfeeding, but the present study shows that these muscles are not essential for the survival of neonatal rats.

Neurotrophic effect of hepatocyte growth factor on neonatal facial motor neurons.

The neurotrophic effect of hepatocyte growth factor (HGF) on axotomized facial motor neurons was examined after local application of HGF to the proximal facial nerve stump of the neonatal rat on post-natal day one (P1). Motor neuron survival was expressed as the neuronal cell count on the injured side as a percentage of that on the noninjured side. Motor neuron survival of the control group was 76% on P3, 54% on P5 and 23% on P8, that of the HGF-treated group 78% on P3, 69% on P5 and 31% on P8, and that of the brain-derived neurotrophic factor (BDNF)-treated group 91% on P5 and 45% on P8. The motor neuron survival rates were then adjusted by deducting the facial motor neurons corresponding to the uninjured retroauricular branch (20%) of the facial nerve. The adjusted values were 70% (P3), 42% (P5) and 4% (P8) for the control group, 72% (P3), 61% (P5) and 14% (P8) for the HGF-treated group, and 88% (P5) and 32% (P8) for the BDNF-treated group. These findings demonstrate that HGF has a neuroprotective effect on injured facial motor neurons and suggest that HGF has neurotrophic properties distinct from those of BDNF.

Proliferating cell populations in experimentally-induced hydrocephalus in developing rats.

To examine the fate of proliferating brain cells in hydrocephalus (Hydro), experimental Hydro was induced in neonatal rats by intracisternal injection of kaolin and, 3 weeks later, the rats were injected with bromodeoxyuridine (BrdU). The BrdU (+) cells were immunohistochemically analyzed by using antibodies against neural (nestin), neuronal (NeuN) and glial (GFAP and MBP) markers in the posterior cerebrum. The percentage of nestin expression for the BrdU (+) cells was 8% in control and increased from 17% in the Hydro to 33% in the Hydro at an earlier stage after the shunt procedure, but was restored to 6% in the Hydro at a later stage after the shunt procedure. The percentages of GFAP expression showed a similar tendency to those of nestin expression. The BrdU (+) cells did not express either NeuN or MBP throughout the experiments.

Great potentiality of neonatal facial motor neurons for neural plasticity as determined by functionally essential neuronal population.

The present study was undertaken to determine the neuronal population essential for normal and minimal facial function of young adult rats that had received various degrees of crush injuries to the facial nerve in the neonatal period. Using a neuronal tracer, it was found in young adult rats receiving neonatal injuries that the minimum number of tracer-labeled facial motor neurons necessary for normal facial function corresponded to 13-14% of the neurons (2540+/-64) of the age-matched control animals, whereas the minimum number of neurons necessary for minimal facial function corresponded to 5%. On the other hand, the minimum numbers of tracer-labeled facial motor neurons necessary for normal and minimal facial function of young adult rats that received various degrees of crush injuries corresponded to 61 and 27-30%, respectively, of the neurons (2540+/-64) of the uninjured control animals. These results indicate that the facial function of animals with nerves crushed at the neonatal stage can be adequately maintained by a very small population of neurons, implying a great potential of neonatal neurons for neural plasticity.

Regenerative process of the facial nerve: rate of regeneration of fibers and their bifurcations.

After the main trunk of the mouse facial nerve was injured by crushing, a fiber tracing method was used to quantify the facial motor neurons that extended regenerating nerve fibers to the specific site of the facial nerve branch. The total number of motor neurons retrogradely labeled with a fluorescent tracer, Fluoro-Gold (FG), were 0 on postsurgical days (PSDs) 1 and 2, 75+/-25 on PSD3, 264+/-21 on PSD4, 378+/-19 on PSD6, 428+/-19 on PSD8, 491+/-13 on PSD12 and 532+/-15 on PSD16. Assuming that the FG-positive neurons (535+/-11) of the control mice represent 100%, the FG-labeled neurons accounted for 0, 14, 49, 71, 80, 92 and 99% on the corresponding days. Two different fluorescent tracers were applied to the different facial nerve branches 16 days after facial nerve injuries. Double-labeled neurons were consistently found in the nerve-crushed facial nucleus (3.2%), and their number increased in the nerve-transected facial nucleus (12.2%). The present study indicates that the regenerating facial nerve consists of heterogeneous nerve fibers with varying growth rates and that excessive axonal branching occurs more frequently in the nerve-transected than in the nerve-crushed injuries.

Functionally essential neuronal population of the facial motor nucleus.

Cranial nerve impairment is one of the more serious complications in neurosurgery. Nevertheless, the important question of how many neurons are required for cranial nerve functions remains unanswered. The VIIth cranial nerve (facial nerve) in mice was subjected to graded crush injuries. After assessment of the facial function, the number of uninjured, healthy facial motor neurons was quantified with a retrograde neuronal tracer. We report that normal facial function is preserved if intact neurons account for more than 56% of the control value, while complete facial paralysis occurs if intact neurons are reduced to less than 32% of the control value.

Differential neurogenesis and gliogenesis by local and migrating neural stem cells in the olfactory bulb.

The rostral migratory stream (RMS) is a unique forebrain structure that provides a long-distance migratory route for the neural stem cells of the periventricular region towards the olfactory bulb (OB). The purpose of the study presented here is to examine the extent of neurogenesis and gliogenesis by the neural stem cells of different origins (periventricular vs. intrabulbar) in the OB. After the RMS had been subjected to injury, the rats received intraperitoneal injections of 5-bromodeoxyuridine (BrdU) and were further reared for 2 weeks. Neuronal and glial differentiations of the BrdU(+) cells in the olfactory bulbar granule cell (OB-GCL) and the olfactory glomerular (OB-GL) layers were examined immunohistochemically using antibodies against neuronal (NeuN, neuronal nuclei) and glial (GFAP, glial fibrillary acidic protein) markers in the OBs with injured and uninjured (control) RMS. In the completely RMS-lesioned OB, where migration of the periventricular neural stem cells was inhibited, a small number of BrdU(+) NeuN(+) cells were found in both the OB-GCL and OB-GL. The BrdU(+) NeuN(+) cells accounted for a much higher percentage of the BrdU(+) cells on the control side (OB-GCL, 36.7%; OB-GL, 8.8%) than on the completely RMS-lesioned side (OB-GCL, 3.7%; OB-GL, 0.6%). The percentage of the BrdU(+) GFAP(+) cells relative to the BrdU(+) cells did not show any major difference between the control and completely RMS-lesioned sides. This study revealed differences in neurogenesis and gliogenesis between the local and migrating neural stem cells in the OB of the adult rodent.

Weak response of helicobacter pylori antibody is high risk for gastric cancer: a cross-sectional study of 10,234 endoscoped Japanese.

BACKGROUND: A large number of endoscoped members of the general Japanese population were surveyed to investigate the relationship between Helicobacter pylori infection and gastric cancer. Special attention was given to antibody titer and age of the subjects. METHODS: We performed gastrointestinal endoscopy and measured serum anti-H. pylori antibody in 10,234 consecutive Japanese who participated in a health examination program. Gastric cancer, when suspected, was confirmed by histology. We graded the H. pylori antibody titer into three groups in accordance with optical density values by ELISA: 'strongly positive', 'weakly positive', and 'negative'. RESULTS: Among the 10,234 subjects (men/women, 7.021/3,213; mean age, 49.1 years), 4,909 (48%) were strongly positive, 1,750 (17%) were weakly positive, and 3,575 (35%) were negative for H. pylori antibody. Thirty-seven cases of gastric cancer were found among the 10,234 subjects (0.36%); 23/4,909 (0.47%) in the strongly positive group, 9/1,750 (0.51%) in the weakly positive group, and 5/3,575 (0.14%) in the negative group. Both the strongly and weakly positive groups showed a higher risk of gastric cancer than the negative group. In the subjects over age 60, the weakly positive group seemed to show the highest risk for gastric cancer. CONCLUSIONS: In this investigation of 10,234 Japanese, based on endoscopy results, those with serum H. pylori antibody had an increased risk for gastric cancer, while those 'weakly positive' showed a high risk, particularly in the elderly.

Cell dynamics of calretinin-immunoreactive neurons in the rostral migratory stream after ibotenate-induced lesions in the forebrain.

It is now apparent that adult neurogenesis is taking place during life in the olfactory bulb (OB) of the rodent brain. In the olfactory nervous system, the precursor cells of the subventricular zone are known to continually proliferate, migrate through the rostral migratory stream (RMS) and differentiate into the bulbar neurons. The RMS, consisting of heterogeneous cell populations of the neural and neuronal precursor cells, is the unique forebrain structure that provides a long-distance migratory route for the precursor cells. The present study was undertaken to examine whether neuronal regeneration, focusing on calretinin-immunoreactive (+) cells, may proceed in the RMS following lesions induced by an excitotoxin. Two days after ibotenate injections, massive degeneration of calretinin (+) cells occurred in the RMS and its adjacent forebrains. Thereafter, calretinin (+) cells gradually increased in the RMS and reached above their control value 2 weeks after ibotenate injections. Removal of the OB also produced a marked increase in calretinin (+) cells in the RMS. Autoradiographic experiments using (3)H-thymidine showed that calretinin (+) cells were continually generated in the RMS and underwent neuronal turnover within 8 weeks in a normal condition. The results indicate that, in terms of calretinin (+) cells, neuronal differentiation and replacement is continually taking place within the RMS, and that the RMS is capable of repopulating those cells which were injured by ibotenate.

The minimum number of neurons in the central olfactory pathway in relation to its function: a retrograde fiber tracing study.

The present study was aimed at determining the functionally essential size of the neuronal population in the central olfactory nervous system. Using conditioned rats who had learnt to avoid repellent (cycloheximide) solution by olfaction, varying degrees of injuries were made to the lateral olfactory tract, a major central olfactory pathway connecting the olfactory bulb to the olfactory cortex. After examining their olfactory ability to discriminate cycloheximide solution from water, intact bulbar projection neurons (mitral cells) with fiber connections to the olfactory cortex were quantified using a retrograde fiber tracing technique. The numbers of retrogradely labeled mitral cells from the rats with normal olfaction ranged between 20 and 92% of the control value, while those numbers from the anosmic rats ranged between 0 and 22%. We conclude that the functionally essential neuronal population is approximately one-fifth of the total in the central olfactory pathway, a presumed threshold value in terms of the ability to avoid cycloheximide solution by olfactory discrimination.

Continual replacement of newly-generated olfactory neurons in adult rats.

It has been known that stem cells do exist in the central nervous system, and adult neurogenesis is continually taking place in the olfactory bulb during life. We report here, with the combined method of autoradiography using (3)H-thymidine and immunohistochemistry for a neuronal marker, that 65.3-76.9% of calretinin-immunoreactive bulbar neurons are replaced during the short period of 6 weeks in the adult rodent. The results indicate that neuronal replacement is a common phenomenon in the olfactory bulb during life.