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C K Govind

Publications and source records attributed to C K Govind.

At least 73 records · Page 4Linked to original sources

Intramembranous organization of lobster excitatory neuromuscular synapses.

The fine structure of identified neuromuscular synapses of the single excitatory axon to the distal accessory flexor muscle in lobster limbs was examined with freeze-fracture and serial thin-section electron microscopy. The latter technique reveals presynaptic dense bars with synaptic vesicles aligned on either side of these bars and often fused to the membrane, suggesting exocytosis and confirming our previous contention that these bars are active zones of transmitter release. The intramembranous organization of these active zones, as revealed in freeze-etched tissue, is a ridge-like elevation of the P-face of the axolemma with a matching trough on the complementary E-face. The ridge on the P-face has rows of large scattered intramembranous particles along the apex and is often bordered by a series of small, circular depressions which are presumed to represent exocytotic vesicles attached to the presynaptic membrane. Complementing these depressions are a few volcano-like protuberances seen occasionally on the E-face membrane. Because such evidence for transmitter release occurred in both stimulated and non-stimulated preparations, it demonstrates that chemical fixatives employing aldehydes induce transmitter release. The postsynaptic receptor sites of these excitatory synapses are characterized by oval-shaped patches of densely packed particles on the E-face, arranged in a random pattern on the sarcolemma. The complementary P-face view exhibits a regular square array of particle imprints or pits.

Aminopyridines↗

Reorganization of synaptic ultrastructure at facilitated lobster neuromuscular terminals.

Prolonged stimulation of the single excitor axon to the lobster distal accessory flexor muscle in the presence of ouabain caused long-term facilitation at its neuromuscular synapses. Hence the extracellularly recorded synaptic potentials failed less frequently and increased their mean amplitude, compared to the non-facilitated (control) potentials from homologous sites in the contralateral muscle. The fine structure of synaptic terminals between matched pairs of facilitated and control preparations was compared with the aid of serial section electron microscopy. Differences between facilitated and control preparations were similar both when the latter were bathed in normal saline or ouabain-containing saline, suggesting that the changes were related to the electrical stimulation rather than to the presence of ouabain. First, the facilitated terminals were smaller in surface area than the control. Second, the number and size of synaptic contacts in the facilitated terminals resembled those in the control. Third, presynaptic dense bodies or active sites increased in number although their sizes remained unaltered in the facilitated terminal. This increase is attributed to the addition of dense bodies at existing synaptic contacts since synaptic contacts remained constant in number between facilitated and control preparations. Fourth, the number and size of synaptic vesicles were unaffected by prolonged stimulation although there was a redistribution of vesicles such that they appeared to be channelled in distinct streams to synaptic contacts. Fifth, mitochondria increased in number and were situated closer to the dense bodies at facilitated nerve terminals than at control terminals. Overall, these changes denote considerable reorganization of the synaptic terminals associated with elevated transmitter release.

Animals↗

Innervation of the limb accessory flexor muscle in several decapod crustaceans. I. Anatomy.

The innervation of the accessory flexor muscle of the limbs of several decapod crustaceans was studied by means of vital staining, with methylene blue and electron microscopy. Three patterns of innervation were found. In the first pattern, the distal (DAFM) and proximal (PAFM) heads of the accessory flexor muscle were supplied by two axons (a thick and a thin) which travel in a private nerve along the length of the merus. This pattern was found in the crab (Cancer) and the lobster (Homarus), and conforms to the classical pattern established in the literature. In the second pattern, the nerve to the DAFM is made up of conjoined branches of the flexor and accessory flexor nerves. Consequently, the DAFM receives at least five axons in the portunid crabs, Carcinus, Callinectes, and Ovalipes, and occasionally six axons in Ovalipes. The PAFM in those portunids receives the usual two axons. In the third pattern, based on preliminary observations on the grapsid crab, Pachygrapsus, "super-innervation" of the accessory flexor muscle appears to include not only five axons to the DAFM but also at least three to the PAFM. In all species, methylene blue staining of the axon terminations revealed a regular pattern of blebs which are thought to correspond to synaptic terminals as revealed by electron microscopy.

Animals↗

Innervation of the limb accessory flexor muscle in several decapod crustaceans. II. Electrophysiology.

The innervation of the distal and proximal heads of the accessory flexor muscle in three portunid crabs and two non-portunid decapods was studied electrophysiologically. In all species studied, the proximal head received only the two previously reported accessory flexor axons, an excitor and an inhibitor. The same two axons also innervated the distal head in all species, but in the portunids the distal head also received excitation from at least three, and probably sometimes four, of the main flexor excitor efferents. The accessory inhibitor exerted very strong effects in the tonic muscle fibers found in the proximal head and in the most proximal bundle of the distal head. The newly described inhibitory and excitatory distributions may have important implications for locomotory behavior.

Action Potentials↗

Enhanced reappearance of fast fibers in regenerating crayfish claw closer muscles.

In the pristine claws of adult crayfish the muscle fibers of the closer are all of slow type as judged by sarcomere lengths of greater than 6 micron, and a uniform degree of myofibrillar ATPase activity. In regenerating claws of mature and immature crayfish, the muscle has a central band of fast type fibers as characterized by shorter sarcomeres (less than 6 micron) and a higher degree of ATPase activity than the surrounding slow fibers. During primary development, the closer muscle has a fiber composition similar to that of the regenerating muscle except for a smaller proportion of fast fibers. Thus the reappearance of fast fibers during regeneration recapitulates ontogeny while their enhanced proportions may reflect epigenetic influences such as restriction of nerve-mediated muscle activity in the limb bud.

Adenosine Triphosphatases↗

Growth-related features of lobster neuromuscular terminals.

Neuromuscular terminals of the low-output type formed by the single excitor axon to the limb distal accessory flexor muscle in the lobster Homarus americanus were studied with serial section electron microscopy. This type of innervation was compared between a small and a large lobster where a two-fold difference in mean quantal content of synaptic transmission was found. Several growth-related changes in the fine structure of these low-output synaptic terminals were seen. First, there was a proliferation of multiterminal innervation consisting of an increase in the number of nerve terminals, synapses and presynaptic dense bars between the small and large lobster. Also the mean surface area of the synapses increased significantly in the large compared to the small lobster. Second, synapses possessed distinct areas of non-specialized membrane or perforations which showed a growth-related increase in their number per synapse between small and large lobsters. Such perforations also occurred in the high-output synapses but only amongst the larger synapses of the older lobster. It is proposed that these perforations subdivided synapses into smaller functional units for membrane recycling as they provide a ready source of non-synaptic axolemma for nearby active sites (dense bars). Third, the branch point between subsidiary and principal terminals as well as the ending of a terminal is composed of synaptic membrane which is presumably involved respectively in the sprouting and elongation of nerve terminals during growth. Altogether these observations signify both qualitative and quantitative changes in identified neuromuscular terminals with growth.

Animals↗

Polyneuronal innervation of an adult and embryonic lobster muscle.

Motor innervation of the deep extensor muscle in the abdomen of lobsters (Homarus americanus) was compared in adults and embryos using electrophysiological techniques. There is widespread innervation of the adult muscle by the common excitor and inhibitor axons and regionally restricted or private innervation by three more excitor axons. In the embryo the earliest sign of functional innervation revealed a single inhibitory and two to three excitatory axons thus denoting simultaneous innervation by the full complement of axons. In corroboration, serial-section electron microscopy revealed several axon profiles invading the embryonic deep extensor muscles and giving rise to well-defined neuromuscular synapses with presynaptic dense bars. Innervation patterns to homologous regions of the embryonic and adult muscles were similar, consisting of a few large inhibitory synapses and many small excitatory ones. Consequently the adult pattern of polyneuronal innervation occurs simultaneously and in toto during embryonic development.

Animals↗

Decrease in transmitter output and synaptic ultrastructure at lobster neuromuscular terminals with decentralization.

The effects of decentralization on the physiology and ultrastructure of neuromuscular terminals were examined by transecting the single excitor axon to the distal accessory flexor muscle in the walking legs of lobsters (Homarus americanus). Decentralization caused a reduction in the amplitude of the excitatory junctional potential without altering the resting potential or input resistance of the muscle fiber thereby suggesting a reduction in transmitter release. Confirmation was obtained by recording of synaptic currents at focal sites which showed failure of transmission and a reduced amplitude on decentralized fibers compared to their intact counterparts on the contralateral leg. The mean quantal content of synaptic transmission decreased approximately 2-7-fold at these decentralized sites compared to their intact counterparts. The ultrastructure of these identified sites was examined with serial section electron microscopy. There are few if any qualitative changes in synaptic ultrastructure between decentralized and control terminals. However, quantitatively there were changes in synaptic ultrastructure which were progressive in nature depending on the severity of the reaction to decentralization. Thus terminals showing a moderate decline in quantal content were characterized by a reduction in the number of presynaptic dense bars and synapses. Terminals showing a severe drop in transmitter release showed in addition to the above changes, a reduction in the size of synapses and terminals. These results show a progression in the loss of the structural parameters controlling transmitter release. Finally synaptic vesicles and mitochondria did not reveal any consistent or marked change with decentralization.

Animals↗

Contractile proteins of fast and slow fibers during differentiation of lobster claw muscle.

Contractile protein populations were determined, using gel electrophoresis, during development of the claw closer muscles of the lobster Homarus americanus. In the adult the paired claw closer muscles are asymmetric, consisting of a crusher muscle with all slow fibers and a cutter muscle with a majority of fast and a few slow fibers. The electrophoretic banding pattern of these adult fast and slow fibers shows a similarity in the major proteins including myosin, actin, and tropomyosin which are common to both fiber types. Paramyosin is slightly heavier in fast fibers than in slow. However, fast fibers have three proteins and slow fibers have four proteins which are unique to themselves. Several of these unique proteins belong to the regulatory troponin complexes. In juvenile 4th stage lobster, where the paired closer muscles are undifferentiated, the banding pattern reveals the presence of proteins common to both fiber types including myosin, actin, and tropomysin but the conspicuous absence of all unique fast fiber proteins as well as one unique slow fiber protein. By the juvenile 10th stage most of these unique proteins are present except for one unique slow fiber protein. Thus lobster fast and slow fiber differentiation entails coordinate gene activation to add unique contractile proteins.

Animals↗

Neural asymmetry in male fiddler crabs.

In adult male fiddler crabs, Uca pugnax, there is a marked enlargement of the 1st thoracic ganglion and its nerve root on the side of the major cheliped compared to the side of the minor cheliped. Retrograde uptake of cobalt via the cut ends of the motoneurons revealed a significant hypertrophy of their somata and dendritic fields on the major side of the ganglion compared to the minor side in the male fiddler crabs. (In female fiddler crabs which have two minor chelipeds the motoneurons were similar in size on both sides of the ganglion.) Since the number and distribution of motoneuron somata was relatively constant in the two halves of each ganglion, homologies for individual or groups of neurons could be recognized. The number of axon profiles in a cross-sectional montage of the entire nerve root of the major side in a male fiddler crab was several times greater than that of the minor side in random samples which were appropriately scaled in area. In samples of equal areas the axonal density was similar on the major and minor sides, as was also the range of axon diameters; both signify no difference in size of axons between the contralateral nerve roots. Consequently enlargement of the nerve root on the major side is due to a relative increase in the number of axons. This increase is in sensory fibers since the number of motor fibers are bilaterally constant. Thus neural asymmetry in male fiddler crabs involves hypertrophy of the motoneurons and hyperplasia of the sensory neurons associated with the enlarged condition of the major cheliped.

Animals↗

Fast and slow motoneurons with unique forms and activity patterns in lobster claws.

The form of the fast closer excitor (FCE) and the slow closer excitor (SCE) motoneurons to the closer muscle in the claw of the lobster Homarus americanus was determined by injecting cobalt chloride or Lucifer Yellow into their respective somata. Both neurons are monopolar with the single neurite rising vertically to the dorsal surface of the ganglion, then travelling along this surface to where it gives off its dendrites before entering the second nerve root as an axon. The FCE and SCE motoneurons, however, differ in their dendritic form in several respects. First, the FCE completely lacks an anterior dendritic field, which is well elaborated in the SCE. Second, the FCE has fewer large primary dendrites in its posterior field than the SCE. Third, the posterior dendritic field of the FCE is not as extensive as that of the SCE. Fourth, the axon of the FCE originates from one of the posterior primary dendrites while that of the SCE is an axial extension of its neurite. Thus the SCE has a more elaborate dendritic field than the FCE, which may account for its greater excitability. For instance, recordings from intact lobsters show that the SCE has a lower firing threshold and is active for longer periods of time and at higher frequencies than the FCE.

Action Potentials↗

Innervation and motor patterns of the abdominal superficial flexor muscles in larval lobsters.

The pattern of innervation and motor program of the abdominal superficial flexor muscle was investigated electrophysiologically in larval lobsters (Homarus americanus). The muscle receives both excitatory and inhibitory innervation in the larval as well as in the embryonic stages. Individual muscle fibers receive a single inhibitory neuron (f5) and a maximum of three excitors. Based on spike heights these axons belong to either the small (f1 or f2) or large (f3, f4) motoneurons. While the small axons preferentially innervate the medial muscle fibers the large axons innervate medial as well as lateral fibers. This larval pattern of innervation resembles the pattern in the adult lobster. The resemblance extends to the firing patterns as well with both large and small excitors firing spontaneously. Furthermore, evoked activity in the larvae produces reciprocal (and occasionally cyclical) bursts of excitor and inhibitor neurons denoting abdominal extension and flexion and resembling the firing patterns in adults. Consequently motor programs employed in steering the pelagic larvae are reminiscent of the programs for maintaining posture in the benthic adult lobsters.

Animals↗

Fine structure of comparable synapses in a mature and larval lobster muscle.

Neuromuscular synapses from the single excitor axon to the proximal accessory flexor muscle (PAFM) was studied by serial section electron microscopy in a 1st stage larval (less than 0.1 g) and a large adult (6.8 kg) lobster. The adult innervation of a lateral and a medial fiber, physiologically identified as low- and high-output respectively, was similar in the number and mean size of synapses but had significantly larger pre-synaptic dense bars for the high-output synapses. This correlation between quantal transmitter output and pre-synaptic dense bars and the appearance of exocytotic profiles along the dense bars strongly implicates the bars as active sites of transmitter release. Moreover the mature innervation is differentiated on the basis that the percentage of dense bar area to synaptic area is 9% for the low-output type compared to 22% for its high-output counterpart. In the larval PAFM the excitatory axon has not proliferated many branches and the innervation is therefore localized to groups of fibers in the lateral, medial and central regions of the muscle rather than to individual fibers. The lateral and medial sites of innervation representing putative low- and high-output types respectively (because of their location) do not differ in the size and number of pre-synaptic dense bars thereby suggesting a similarity in quantal synaptic transmission. However, the percentage of dense bar area to synaptic area is 40% for the lateral site compared to 67% for the medial site. Since this is a trend mimicking the mature innervation it shows an early stage in the differentiation of low- and high-output synapses. Furthermore the main axon provides half of the total innervation in the larval PAFM but none in the adult thereby demonstrating a restructuring of multiterminal innervation.

Animals↗

Differentiation of identifiable lobster neuromuscular synapses during development.

The ultrastructure of physiologically identified low and high release synapses arising from a single axon on fibres of the distal accessory flexor muscle (DAFM) in a mature lobster was examined by serial section electron microscopy. Low release neuromuscular terminals located only on the proximal fibre were characterized by large synapses (mean area 2.084 micron2), small presynaptic dense bars (mean are 0.021 micron2) and hence a low (2.3%) ratio of dense bar area to synaptic area. In contrast high output terminals located only on the distal fibre had smaller synapses (mean area 0.625 micron2), large dense bars (mean area 0.066 micron2) and a high (23.9%) ratio of bar area to synaptic area. A similar ratio was consistently found for each synaptic type in several other examples of mature lobsters. Hence it was used as a criterion for determining the point at which differentiation occurs during development. In the first larval stage (24 h old) the innervation was localized and undifferentiated. In the fourth (2 week old) and twelfth (1 y old) stage lobsters, the innervation had proliferated to small bundles of proximal and distal fibres. During development synapses increase in their mean surface area in the proximal fibre while remaining constant in the distal fibre. The mean surface area of the dense bars is similar in all stages except for the proximal fibres of the twelfth stage where it is smaller by 50%. Similarly the ratio fo dense bar area to synaptic area is not significantly different for all stages except for the twelfth stage proximal fibres where it is half the value. Consequently differentiation of low and high release neuromuscular terminals occurs by the twelfth stage with an increase in the mean surface area of synapses and a decrease in the mean surface area of dense bars. This morphological differentiation is enhanced in the mature lobster.

Animals↗

Arrays of particles in the sarcolemma of the distal accessory flexor muscle of the lobster.

Freeze-fracture of the distal accessory flexor muscle reveals the presence of randomly distributed arrays of P- and E-face particles. The particles are arranged in regularly spaced rows which parallel one another and, in general, the long axis of the muscle fiber. P- and E-face arrays differ in their structure and are apparently complementary. A model explaining the apparent interrelationship of the P- and E-face arrays is presented. The functional significance of the arrays is not known.

Animals↗