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Motoharu Hayashi

Publications and source records attributed to Motoharu Hayashi.

27 records · Page 2Linked to original sources

Levels of serum brain-derived neurotrophic factor in primates.

Brain-derived neurotrophic factor (BDNF) has been reported to exist not only in nervous tissue but also in serum. In contrast to the wealth of knowledge regarding the various physiological functions of BDNF in the nervous system, information about possible roles in other systems is limited. To elucidate the physiological function of serum BDNF in primates, it is first necessary to establish a method to determine the levels of BDNF in serum of primates. In the present study, we established an enzyme-linked immunosorbent assay (ELISA) method which we used to measure levels of serum BDNF in non-human primates. We found that serum BDNF levels were similar among several species of primates. The present results suggest that our BDNF ELISA may be useful in measuring serum BDNF concentration as a physiological marker, and that levels of serum BDNF may be similar among primates including humans.

Animals↗

Comparative study of urinary reproductive hormones in great apes.

Urinary estrone conjugates (E(1)C), pregnanediol-3-glucuronide (PdG), and follicle-stimulating hormone (FSH) were determined by enzyme immunoassays (EIAs) during the normal menstrual cycle in the orangutan, gorilla, chimpanzee, and bonobo. Furthermore, the data were compared to those levels in the human and long-tailed macaque. The results showed a typical preovulatory E(1)C surge and postovulatory increase in PdG in all species. The pattern of E(1)C during the menstrual cycle in the great apes more closely resembled the human than do the long-tailed macaque. A major difference of E(1)C pattern between these species appeared in the luteal phase. In the great apes and the human, E(1)C exhibited two peaks, the first peak detected at approximately mid cycle and the second peak detected during the luteal phase. On the other hand, in the long-tailed macaque, increase of E(1)C in the luteal phase was small or nonexistent. The gorilla, chimpanzee, and bonobo exhibited similar PdG trends. The orangutan excreted one tenth less PdG than these species during the luteal phase. The long-tailed macaque also excreted low levels of PdG. The patterns of FSH in orangutan, chimpanzee, bonobo and long-tailed macaque showed a marked mid-cycle rise and an early follicular phase rise, similar to those in the human. Comparing similar taxa, a large difference was found in FSH of gorilla; there were three peaks during the menstrual cycle. Thus, there is considerable species variation in the excretion of these hormones during the menstrual cycle and comparative studies could be approached with a single method. The methods and baseline data presented here provide the basis for a practical approach to evaluation and monitoring of ovarian events in the female great apes.

Animals↗

Expression of TrkB subtypes in the adult monkey cerebellar cortex.

BDNF and its specific receptor TrkB are concerned with synaptic plasticity as well as maintenance of the nervous system. TrkB has three subtypes: full-length TrkB (TK+), which has a tyrosine kinase containing intracellular domain, and two truncated TrkBs (TK-; T1 and T2), which lack tyrosine kinases. To understand the molecular interaction among these subtypes, we investigated the expression and distribution of BDNF, TK+, and T1 in the adult monkey cerebellum by single and double immunohistochemistry and Western blot analysis. We observed by single immunohistochemistry that BDNF, TK+, and T1 are distributed in almost all the somata and dendrites of Purkinje and granule cells. In the double-stained sections, three kinds of regions were observed: TK+ >T1; TK+ =T1; TK+ <T1. Moreover, three types of TrkB dimers (TK+/TK+ homodimer, TK+/TK- heterodimer, and TK-/TK- homodimer) were induced by stimulating with exogenous BDNF. These observations suggest that the functions of BDNF may be modified by interaction among subtypes of TrkB in each region of the Purkinje cells.

Animals↗

Urinary steroids, FSH and CG measurements for monitoring the ovarian cycle and pregnancy in the chimpanzee.

Non-invasive methods for monitoring reproductive status of chimpanzee based on the measurement of urinary steroids and gonadotropins were examined. A typical pre-ovulatory urinary estrone conjugate (E1C) surge and post-ovulatory increase in pregnandiol glucuronide (PdG) were seen during the menstrual cycle. Urinary follicle stimulating hormone (FSH) showed two peaks over the infertile menstrual cycle. The earliest changes indicating pregnancy were a coincident rise in E1C and chorionic gonadotropin (CG) levels and a concomitant fall in FSH levels. Urinary PdG levels showed a prolonged rise. Urinary E1C in the pregnant chimpanzee was higher than during the menstrual cycle and increased with advancing gestation, with maximum levels occurring near term. In the case of stillbirth, E1C and CG levels from mid- through late-pregnancy were low and the prepartum progressive increase in E1C was not shown. The data presented here are of great practical value in captive breeding management of chimpanzees.

Animals↗

Comparison of mineral contents between the arteries in upper and lower limbs of Japanese monkeys.

To examine whether the calcium accumulation in aged arteries is related to the way of walking, the mineral contents were determined in the arteries of Japanese monkeys of quadrupedal walk by inductively coupled plasma-atomic emission spectrometry. Sixteen Japanese monkeys consisting of 7 males and 9 females ranging in age from 2 to 33 yr were studied. The accumulation of calcium, phosphorus, and magnesium occurred progressively in most, but not all, of the arteries with aging. It was found that independent of the upper and lower limbs, a higher accumulation of calcium, phosphorus, and magnesium occurred in the arteries of the proximal regions with aging, compared with the arteries of the distal regions. In a comparison between the arteries of anatomically corresponding regions of the upper and lower limbs, the accumulation of calcium and magnesium was 20-60% higher in the external iliac and femoral arteries of the lower limb than in the axillary and brachial arteries of the upper limb. Regarding phosphorus, the accumulation was 20-120% higher in the external iliac and femoral arteries than in the axillary and brachial arteries. It was known that in humans, the accumulation of calcium, phosphorus, and magnesium was three to seven times higher in the arteries of the lower limb than in the arteries of the upper limb. It is clear that there is a very significant difference in the accumulation of calcium and magnesium in the arteries of the lower limbs between Japanese monkeys and humans. The present study suggests that the accumulation of calcium and magnesium in the arteries of the lower limb with aging is affected by the way of walking.

Aging↗

Changes in NT-3 and TrkC in the primary visual cortex of developing macaques.

In the present study, we measured the levels of neurotrophin-3 (NT-3) protein and mRNA by ELISA and quantitative RT-PCR, and examined TrkC-immunoreactive structures using immunohistochemical method. In the nervous system of the adult monkey, higher levels of NT-3 were found in the hippocampus and cerebellum. During the development of the primary visual cortex, detected amounts of NT-3 protein peaked at embryonic day 140 and then gradually decreased. TrkC imunoreactivity was observed in the neurons in layer VI of the primary visual cortex of an embryonic monkey. These results support the hypothesis that NT-3 is involved in the specification of axon targeting from layer VI to layer IV during the late embryonic stages.

Aging↗

Expression of super cervical ganglion-10 (SCG-10) mRNA in the monkey cerebral cortex during postnatal development.

We used Northern blot analysis to measure SCG10 mRNA expression in various areas of the monkey cerebral cortex during postnatal development. In all areas, SCG10 mRNA expression was highest immediately after birth, decreased sharply until postnatal day 70 (P70) and then decreased slowly until the adult stage. Expression levels were similar in all areas at all time points during development, including the adult stage. These findings confirm that axonal growth in the monkey cerebral cortex is most vigorous before or during the perinatal period, progresses similarly in all areas until P70, and continues at lower levels during late development.

Aging↗

Northern blot and in situ hybridization analyses of MARCKS mRNA expression in the cerebral cortex of the macaque monkey.

Myristoylated alanine-rich C-kinase substrate (MARCKS) is a major substrate for protein kinase C, and is involved in synaptic plasticity. Using both Northern blot and in situ hybridization techniques, we investigated whether MARCKS expression varied according to the cerebral region, including the hippocampal formation, or according to the type of neuron. Northern blot analysis showed that the MARCKS mRNA level was higher in the association areas than in the primary sensory and motor areas of the cerebral neocortex. MARCKS mRNA levels in the hippocampus and the amygdala were as high as those in the association areas. The in situ hybridization experiments confirmed the Northern blot results and showed the distribution and characteristics of MARCKS mRNA-positive neurons. In the association areas of the neocortex, prominent signals were observed in neurons in layers II-VI. In the primary areas, prominent signals were restricted to neurons in layers IV-VI. In the hippocampus, the most intense hybridization signals were observed in neurons in the granule cell layer of the dentate gyrus. The observed region-specific expression might reflect functional specialization for plasticity in individual regions of the monkey cerebral cortex.

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Expression of MARCKS mRNA in lateral geniculate nucleus and visual cortex of normal and monocularly deprived macaque monkeys.

We performed a nonradioactive in situ hybridization histochemistry (ISH) study of the lateral geniculate nucleus (LGN) and the primary visual area (area 17) of the macaque monkey to investigate mRNA expression of the myristoylated alanine-rich C-kinase substrate (MARCKS), a major protein kinase C (PKC) substrate. In the LGN, intense hybridization signals were observed in both magnocellular neurons (layers 1 and 2) and parvocellular neurons (layers 3 to 6). Double labeling using ISH and immunofluorescence revealed that MARCKS mRNA was coexpressed with the alpha-subunit of type II calcium/calmodulin-dependent protein kinase, indicating that MARCKS mRNA is also expressed in koniocellular neurons in the LGN. GABA-immunoreactive neurons in the LGN did not contain MARCKS mRNA, indicating that MARCKS mRNA is not expressed in inhibitory interneurons. The signals were generally weak in area 17, and intense signals were restricted to large neurons in layers IVB, V, and VI. GABA-immunoreactive neurons in layers II-VI of area 17 did not contain MARCKS mRNA. Double-label ISH revealed that MARCKS mRNA was coexpressed with mRNA of GAP-43, another PKC substrate, in neurons of both the LGN and area 17. To determine whether the expression of MARCKS mRNA is regulated by retinal activity, we performed ISH in the LGN and area 17 of monkeys deprived of monocular visual input by tetrodotoxin. After monocular deprivation for 5 to 30 days, MARCKS mRNA was down-regulated in the LGN, but not in area 17. These results suggest that MARCKS mediates the activity-dependent changes in the excitatory relay neurons in the LGN.

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