Quantitative analysis of development and aging of genital corpuscles in glans penis of the rat.

The aim of the present postnatal developmental study was to determine densities of unique genital corpuscles (GCs) in glans penis of developing and aged rats. GCs were identified as corpuscular endings consisting of highly branched and coiled axons with many varicosities, which were immunoreactive for protein gene product 9.5. In addition, GCs were immunoreactive for calcitonin gene-related peptide and substance P, but not for vasoactive intestinal polypeptide and neuropeptide Y. GCs were not found in the glans penis of 1 week old rats. Densities of GCs were low at 3 weeks, significantly increased at 5 and 10 weeks, reached the peak of density at 40 weeks, and tended to decrease at 70 and 100 weeks. Sizes of GCs were small in 3 weeks old rats, increased at 5 and 10 weeks, reached the peak-size at 40 weeks and reduced in size at 70 and 100 weeks. Considering sexual maturation of the rat, the results reveal that GCs of the rat begins to develop postnatal and reaches to the peak of their development after puberty and continues to exist until old age, in contrast to prenatal and early postnatal development of other sensory receptors of glabrous skin.

Evolutionary anatomy and phyletic implication of the extrinsic cardiac nervous system in the Philippine tarsier (Tarsius syrichta, Primates) in comparisons with Strepsirrhines and New World monkeys.

To provide information on evolutionary morphological differences and similarities between Tarsiformes (tarsiers) and other primates (Strepsirrhini and Platyrrhini), we examined the previously undescribed extrinsic cardiac nervous system (ECNS) and surrounding structures in three Philippine tarsiers (Tarsius syrichta). We found that the complicated ECNS morphology shows little variation in contrast to significant anatomical diversity of the surrounding somatic structures in tarsiers as reported in our previous primate studies. Further, our comparative morphological data on the ECNS show the following aspects of evolutionary anatomical significance; (1) the traits of the general relationship between the superior cervical ganglion and the spinal nerves are common to the Tarsii and Strepsirrhini, (2) the traits of the general composition of the cervicothoracic ganglion, its communicating branches to spinal nerves, and its cardiac nerve are common to the Tarsii and Platyrrhini, and (3) the traits of the general relationship between the middle cervical ganglion and the spinal nerves and its cardiac nerve are common among Strepsirrhini, Tarsii, and Platyrrhini. Therefore, the general anatomical characteristics of the ECNS in tarsiers retain a primitive state for some traits and exhibit a derived state for others. These characteristics of the ECNS also reflect an evolutionary history without the modification and specialization as seen in the surrounding somatic structures. It may play a key role in unifying future studies on molecular phylogeny and updating evolutionary anatomical information.

Triadic synaptic interactions of large corticothalamic terminals in non-lemniscal thalamic nuclei of the cat auditory system.

Large corticothalamic (CT) terminals, presumed to originate from cortical layer 5 pyramidal cells, are distributed predominantly in non-specific thalamic nuclei in mammals. In the auditory system, little is known about whether these CT projections participate in the synaptic aggregation referred to as the triad. We studied synaptic interactions of these terminals with neuronal elements in one of the auditory non-lemniscal thalamic nuclei, the dorsal nucleus of the medial geniculate complex (MGC), in cats. After injections of an anterograde tracer in the primary auditory cortex, areas containing labeled large terminals were examined using an electron microscope. It was revealed that a fraction of large CT terminals participated in complicated synaptic arrangements: labeled terminals making synaptic contacts with vesicle-free dendrites, probably of thalamic principal neurons, and/or vesicle-filled neuronal profiles, probably of presynaptic dendrites (PSDs) of interneurons. In reconstructions or even in single sections, we found that these synaptic connections participated in triadic arrangements. Thus, PSDs postsynaptic to the labeled CT terminals were in turn presynaptic to the vesicle-free dendrites.

[Attempt to reduce the formaldehyde concentration by blowing cooled fresh air down in to the breathing zone of medical students from an admission port on the ceiling during gross anatomy class].

Cadavers in gross anatomy laboratories at most medical schools are conventionally embalmed in formaldehyde solution, which is carcinogenic to humans. Medical students and instructors are thus exposed to formaldehyde vapors emitted from cadavers during dissection. To reduce high formaldehyde concentrations in the breathing zone above cadavers being examined by anatomy medical students provisionally, dissection beds were located under existing admission ports on the ceiling to supply cooled fresh air from the admission port blowing downward on to the cadaver. In all cases, compared to normal condition, the downward flow of cooled fresh air from an admission port reduced formaldehyde concentrations by 0.09-0.98 ppm and reduced to 12.6-65.4% in the air above a cadaver in the breathing zone of students. The formaldehyde concentrations above cadavers under admission ports were not more than the formaldehyde concentrations between beds representing the indoor formaldehyde concentrations. Although the application of an existing admission port on the ceiling in this study did not remove formaldehyde, the downflow of cooled fresh air using this system reduced the formaldehyde concentration in the air above cadavers being attended by anatomy students during dissections. These results suggest the need for reducing formaldehyde levels in gross anatomy laboratories using fundamental countermeasures in order to satisfy the guidelines of 0.08 ppm established by the World Health Organization and the Japan Ministry of Health, Labor and Welfare.

[Vertical distribution of formaldehyde concentration and simulated temperature and wind velocity from computational fluid dynamics in a gross anatomy laboratory].

Cadavers for gross anatomy laboratories are typically embalmed in formaldehyde. Thus, medical students and instructors are exposed to formaldehyde vapors emitted from cadavers during dissection. In an attempt to improve the dissection environment, we examined indoor formaldehyde concentrations in a gross anatomy laboratory. Air samples were taken from 20, 110, 160, and 230 cm above the floor between dissection beds to represent areas near the floor, in the breathing zone of sitting students, in the breathing zone of standing students, and near the ceiling, respectively. Formaldehyde vapors were thoroughly diffused from the floor to the ceiling, suggesting that medical students are exposed to similar concentrations of formaldehyde based on distance from the floor. Computational fluid dynamics showed that cadavers are warmed by overhead fluorescent lights and the body heat of anatomy students, and indicated that the diffusion of formaldehyde vapors is increased by lighting and the body temperature of students. Computational fluid dynamics showed that gentle convection from anatomy students and cadavers carry formaldehyde vapors upward; downward flow near admission ports diffuse formaldehyde vapors from the ceiling to the floor in the anatomy laboratory.

Innervation of the triangular fibrocartilage complex of the human wrist: quantitative immunohistochemical study.

The distribution of neural elements in the triangular fibrocartilage complex (TFCC) of the human wrists was studied via immunohistochemical staining of protein gene product (PGP) 9.5 and calcitonin gene-related peptide (CGRP). Articular branches projecting to the TFCC arose from the dorsal branch of the ulnar nerve in all wrists examined. The TFCC is subdivided into the following six regions: the articular disc proper (ADP), meniscus homolog (MH), radio-ulnar ligament (RUL), loose part of ulnar collateral ligament (lUCL), dense part of ulnar collateral ligament (dUCL), and internal portion (IP). The IP consists of a mixture of dense and loose connective tissues enclosed by the ADP, MH, RUL, and UCL, and resides deep in the prestyloid recess, which is a pit in the MH. The densities of PGP 9.5-positive neural elements, including free nerve endings, single nerve fibers, nerve fascicles, and perivascular neural nets, were significantly higher in the IP than in other regions. Some of the neural elements except for the perivascular neural nets were positive for CGRP. The high density of neural elements in the IP suggests that sensory nerves projecting to the TFCC enter into the IP and from there distribute to adjacent regions such as the MH and RUL. Free nerve endings are responsible for pain transmission. The high density of free nerve endings in the IP suggests that the IP is a source of ulnar side wrist pain.

[Formaldehyde concentrations in the breathing zone of medical students during gross anatomy laboratory in Toho University].

Cadavers for gross anatomy laboratories are conventionally embalmed by formaldehyde (FA) solution in most medical schools. Thus, medical students and instructors are exposed to FA vapors emitted from cadavers during dissection. As a basic survey for the improvement of the dissection environment, we examined FA concentration in the gross anatomy laboratory during the 2006 academic year at the Faculty of Medicine of Toho University. Air samples were taken from 20 cm above a cadaver as breathing zone, and above a desk between cadavers as indoor FA concentration. FA concentrations in the breathing zone were ranged from 0.24 to 3.04 (mean 1.71) ppm during systematic anatomy, and from 0.72 to 1.60 (mean 1.16) ppm during neuroanatomy, and indoor FA concentration ranged from 048 to 1.11 (mean 0.76) ppm and from 0.21 to 0.23 (mean 0.22) ppm, respectively. These results showed that medical students and instructors are exposed to higher concentrations of FA than allowed by the guidelines of the Japan Ministry of Health, Labor and Welfare, and suggested the need to reduce FA levels in the gross anatomy laboratory.

Quantitative analysis of axon collaterals of single superficial pyramidal cells in layer IIb of the piriform cortex of the guinea pig.

To understand the functional organization of the piriform cortex (PC), the axon collaterals of three pyramidal cells in layer IIb of the anterior PC and one pyramidal cell in layer IIb of the posterior PC were labeled and quantitatively analyzed by intracellular biocytin injection in the guinea pig. Single pyramidal cells in the anterior and posterior PCs have widely distributed axon collaterals, which exhibit little tendency for patchy concentrations inside as well as outside the PC. The total lengths of the axon collaterals of the three fully analyzed pyramidal cells ranged from 68 to 156 mm, more than 50% of which were distributed in the PC. The total number of boutons of the three cells ranged from 6000 to 14,000, 5000-7000 of which were distributed in the PC. It was estimated that individual pyramidal cells in layer IIb form synaptic contacts with 2200 to 3000 other pyramidal cells in the PC, indicating that single pyramidal cells in layer IIb receive input from a large number of other pyramidal cells. This high connectivity of the network of pyramidal cells in the PC can be regarded as the neural network operating parallel distributed processing, which may play an important role in experience-induced enhancement in odorant discrimination in the PC.

Quantitative analysis of axon collaterals of single neurons in layer IIa of the piriform cortex of the guinea pig.

To study the various types of neurons in layer IIa in the piriform cortex (PC) and the spatial distribution of their axons, axon collaterals of three neurons in layer IIa were labeled and quantitatively analyzed by intracellular injection of biocytin in the guinea pig. Individual neurons have highly distributed axon collaterals, which display a little tendency toward patchy concentrations inside as well as outside the PC. One semilunar cell in the posterior PC had 54-mm-long axon collaterals and 4,200 boutons, out of which 2,100 (49% of the total number of boutons) were distributed in the PC. One semilunar-pyramidal transitional cell in the posterior PC had 256-mm-long axon collaterals and 23,000 boutons, out of which 16,100 (70% of the total number of boutons) and 4,000 (18% of the total number of boutons) were respectively distributed in all layers and in layer Ia of the PC. One multipolar cell in the posterior PC had 188-mm-long axon collaterals and 18,000 boutons, out of which 13,700 (78% of the total number of boutons) were distributed in the PC. Our results revealed that the connection patterns of individual cells in layer IIa have most of the features required for an associative neural network, which may function as a content-addressable memory for the association of odor stimuli.

Quantitative analysis of axon collaterals of single cells in layer III of the piriform cortex of the guinea pig.

Recent physiological and morphological studies suggest that the piriform cortex (PC) functions like the association areas of the neocortex rather than the typical primary sensory area as was previously assumed. The axon connection patterns of single cells are important for understanding the functional organization of the PC. The axon collaterals of three single pyramidal cells and one spiny multipolar cell in layer III of the PC were labeled and quantitatively analyzed by intracellular injections of biocytin in guinea pigs. The individual pyramidal and spiny multipolar cells have highly distributed axon collaterals, which display little tendency for patchy concentrations, within the PC and multiple higher order behavior/reward/contextual-related areas, such as the prefrontal cortex, amygdaloid nuclei, and entorhinal cortex. For the pyramidal cells, the average length of axonal collaterals is 143 mm; the average number of boutons is 12,930. For the spiny multipolar cell, the length of the axonal collaterals is 88 mm; the number of boutons is 7,052. The pyramidal cells in the anterior subdivision of the PC (APC) have both rostrally and caudally directed intrinsic association fibers, whereas the pyramidal and spiny multipolar cells in the posterior subdivision (PPC) have predominantly caudally directed intrinsic association fibers in the PC. Our results reveal that the connection patterns of single cells in layer III resemble those of pyramidal cells in layer II, suggesting that the PC performs correlative functions analogous to those in the association area of other sensory systems. The rostrally-to-caudally directed connections in the APC provide a substrate for the recurrent process, whereas largely caudally directed connections in the PPC suggest the dominance of the feed-forward process.

Microglial ecto-Ca(2+)-ATPase activity in a rat model of focal homologous blood clot embolic cerebral ischemia: an enzyme histochemical study.

Post-ischemic changes in ecto-Ca(2+)-ATPase activity in microglia and the infarcted tissue were studied in a rat model of focal embolic cerebral ischemia using an enzyme histochemical method. Ecto-Ca(2+)-ATPase activity was observed in whole brains in non-operated and sham-operated control animals. In addition, this enzyme activity was determined to be localized in ramified microglia. At 30 min after ischemia, non-microglial ecto-Ca(2+)-ATPase activity in the infarcted tissue slightly decreased and continued to decrease thereafter. The ecto-Ca(2+)-ATPase activity in microglia did not appear changed at this time. The decrease of enzyme activity in the infarcted tissue made it much easier to clearly observe ecto-Ca(2+)-ATPase-positive microglia. The enzyme activity of microglia in the ischemic area began to decrease 2 or 4h after embolization and remarkably decreased, except in the perinuclear cytoplasm, apical parts of the processes, and several parts along the processes, 8h after ischemia. By 12h after onset of embolization, the enzyme activity of microglia and infarcted tissue had almost completely disappeared. Ecto-Ca(2+)-ATPase of microglia is likely to play an important role in the metabolism of extracellular nucleotides in the ischemic area immediately after the onset of embolization by means of ecto-enzymes. Thus, the findings of the present study suggest that microglia might serve to protect the infarcted tissue in the ischemic brain.

Intracellular characterization of suppressive responses in supragranular pyramidal neurons of cat primary auditory cortex in vivo.

Several suppressive processes shape the response properties of auditory neurons, namely lateral inhibition, non-monotonic rate level function and excitation/inhibition binaural interaction. By combining intracellular recording from and staining of layers 2 and 3 pyramidal neurons (PNs) in cat primary auditory cortex, we demonstrate the temporal aspects of depolarization and hyperpolarization underlying these suppressions using pure tone stimulation. Two populations can be distinguished by the occurrence of hyperpolarization following onset depolarization (O-DEP). In layer 2 PNs there is an absence of hyperpolarization following O-DEP, while in layer 3 PNs hyperpolarization follows O-DEP. The latency of O-DEP is shortest at the neuron's best frequency. The latency shortens as sound intensity increases. In non-monotonic PNs, hyperpolarization onset becomes shorter as sound intensity increases. This earlier onset of hyperpolarization shortens the duration of the preceding O-DEP, resulting in a decreased O-DEP amplitude. Diverse patterns in the temporal interaction of depolarization and hyperpolarization underlie the binaural suppression interaction. These results demonstrate that diverse suppressive responses result from differences in the temporal timing of excitation and inhibition. The present results also suggest the possibility of distinct connections between PNs responding in a similar manner.