Biomedical subjects

R H Anken

Publications and source records attributed to R H Anken.

At least 37 records · Page 2Linked to original sources

PubMed2001

Vesicular bodies in fish maculae are artifacts not contributing to otolith growth.

The presence, morphology and possible origin of vesicle-like bodies (VBs) within the inner ear macular otolithic membrane of developmental stages of cichlid fish Oreochromis mossambicus and neonate (i.e. functionally fully developed except the reproductive organs) swordtail fish Xiphophorus helleri were analyzed by means of transmission and scanning electron microscopy (TEM and SEM, respectively) employing various fixation procedures. Some authors believe that these VBs are involved in the formation of the organic phase of inner ear otoliths (or statoliths in birds and mammals). Decreasing the osmolarity of the fixation medium from a value rather close to that of native fresh water fish tissue (i.e. 250 mOsm and 290--300 mOsm, respectively) to a value of fixatives mostly employed in TEM studies (ca. 190 mOsm), the amount of VBs increased and the components of sensory inner ear tissue increasingly dilated. Whilst a conventional prefixation with aldehydes followed by osmium tetroxide postfixation yielded numerous VBs, only few of them were observed when the tissue was fixed with aldehydes and osmium tetroxide simultaneously. Therefore, the results demonstrate that inner ear sensory epithelia are extremely sensitive to altering fixation media. On this background it must be concluded that VBs are fixative (i.e. glutaraldehyde) induced artificial structures, so-called membrane blisters. Thus, the protein matrix of otoliths (and possibly that of statoliths in higher vertebrates) is rather provided by secretion processes than by the release of vesicles.

Acoustic Maculae↗

PubMed2001

Effect of hypergravity on the Ca/Sr composition of developing otoliths of larval cichlid fish (Oreochromis mossambicus).

The amounts of calcium and strontium were measured by inductively coupled plasma mass spectrometry (ICP-MS) in saccular and utricular inner ear otoliths (sagittae and lapilli, respectively) of developing cichlid fish. These fish had been maintained for 22 days at 3-g hypergravity conditions within a centrifuge. During this time-span, the animals completed their ontogenetic development from hatch to the free-swimming stage. Neither the morphogenetic development nor the timely onset and gain of performance of the swimming behaviour was impaired by the experimental conditions. Experimental and control animals also did not differ concerning their size (total length). ICP-MS revealed that the otoliths contained significantly less calcium (in microg/otolith) after hyper-g exposure compared to parallelly raised 1-g control specimens (lapilli: 0.74+/-0.21 vs. 1.16+/-0.41; sagittae: 2.09+/-0.49 vs. 2.76+/-0.47). The content of strontium (in microg/otolith: lapilli: 0.0044+/-0.0023 vs. 0.0022+/-0.0013; sagittae: 0.0094+/-0.0026 vs. 0.0081+/-0.0016) and, consequently, the Sr/Ca ratio (Sr/Cax100) was increased (lapilli: 0.607+/-0.267 vs. 0.201+/-0.12; sagittae: 0.439+/-0.093 vs. 0.301+/-0.086). Since the calcium content can be taken as a proxy for otolith weight, and because parallelly undertaken morphometric investigations revealed smaller otoliths (maximum radius and surface area) due to hyper-g exposure, the results suggest that the growth of otoliths at hyper-g is slowed down. Since the concentration of trace elements incorporated into otoliths is likely based on the composition of the respective protein matrix, our findings suggest that the protein metabolism is affected by hypergravity.

Animals↗

PubMed2000

Weightlessness during spaceflight results in enhanced synapse formation in a fish brain vestibular nucleus.

Synapse counts were undertaken by conventional electron microscopy in primary vestibular integration centers, (i.e. nucleus descendens and nucleus magnocellularis of the brainstem area octavolateralis) and in the diencephalic visual nucleus corticalis of spaceflown neonate swordtail fish Xiphophorus helleri as well as in 1 g control siblings. Spaceflight (16 days microgravity, (microg), STS-90 Neurolab Mission) yielded an increase in synaptic contacts within the vestibular nucleus descendens indicating that lack of input resulted in compensation processes. No effect of microg, however, was observed in the visual nucleus corticalis and in the vestibular nucleus magnocellularis which is situated in the close vicinity of the nucleus descendens. In contrast to the latter, the nucleus magnocellularis does not receive exclusively vestibular input, but inputs from the lateral line as well, possibly providing sufficient input at microgravity.

Animals↗

PubMed2000

Fish inner ear otoliths stop calcium incorporation after vestibular nerve transection.

Previous investigations revealed that the growth of fish inner ear otoliths (otolith size and calcium incorporation) depends on the amplitude and the direction of gravity, suggesting the existence of a (negative) feedback mechanism. In a search for the regulating unit, the vestibular nerve was unilaterally transected in neonatal swordtail fish (Xiphophorus helleri) which were subsequently incubated in the calcium-tracer alizarin-complexone. Calcium incorporation and thus otolith growth ceased on the operated head sides, indicating that the brain is significantly involved in regulating otolith growth.

Animals↗

PubMed2000

Gravitational neurobiology of fish.

In vertebrates (including man), altered gravitational environments such as weightlessness can induce malfunctions of the inner ears, based on irregular movements of the semicircular cristae or on dislocations of the inner ear otoliths from the corresponding sensory epithelia. This will lead to illusionary tilts, since the vestibular inputs are not confirmed by the other sensory organs, which results in an intersensory conflict. Vertebrates in orbit therefore face severe orientation problems. In humans, the intersensory conflict may additionally lead to a malaise, commonly referred to as space motion sickness (SMS), a kinetosis. During the first days at weightlessness, the orientation problems (and SMS) disappear, since the brain develops a new compensatory interpretation of the available sensory data. The present review reports on the neurobiological responses--particularly of fish--observed at altered gravitational states, concerning behaviour and neuroplastic reactivities. Recent investigations employing microgravity (spaceflight, parabolic aircraft flights, clinostat) and hyper-gravity (laboratory centrifuges as ground based research tools) yielded clues and insights into the understanding of the respective basic phenomena.

Animals↗

PubMed2000

Readaptation of fish to 1g after long-term microgravity: behavioural results from the STS 89 mission.

The swimming behaviour of adult and neonate swordtail fish Xiphophorus helleri was qualitatively analysed from video recordings taken throughout the STS 89 spaceshuttle mission from launch to landing and thereafter. After the flight, the swimming behaviour of neonate samples was quantitatively assessed in the course of the readaptation to 1g earth gravity at days 0, 1 and 4 after recovery. Regarding the swimming behaviour during the mission, the adult fish swam thigmotactically (i.e., responding to tactile stimuli) along the walls of their aquarium, but like the neonates, they did not show any aberrant behavioural patterns. This indicates that they could easily adapt themselves to microgravity. On mission day 9, however, looping responses (most probably initiated by mechanical disturbances) occurred indicating a continuously performed "C-start" escape response (the respective body bend looks like the letter "C"). Immediately after landing (observed in videos recorded onboard the space shuttle), the adults performed a head-up swimming beating heavily with the caudal and pectoral fins; this aberrant behaviour gradually decreased during the first hours after recovery.

Adaptation, Physiological↗

PubMed2000

Fish otolith growth in 1g and 3g depends on the gravity vector.

Size and asymmetry (size difference between the left and the right side) as well as calcium (Ca) content of inner ear otoliths of larval cichlid fish Oreochromis mossambicus were determined after a long-term stay at hypergravity conditions (3g; centrifuge). Both utricular and saccular otoliths (lapilli and sagittae, respectively) were significantly smaller after hyper-g exposure as compared to parallely raised 1g-control specimens and the absolute amount of otolith-Ca was diminished. The asymmetry of sagittae was significantly increased in the experimental animals, whereas the respective asymmetry concerning lapilli was markedly decreased. In the course of another experiment larvae were raised in aquarium hatch baskets, from which one was placed directly above aeration equipment which resulted in random water circulation shifting the fish around ("shifted" specimens). The lapillar asymmetry of the "stationary" specimens showed a highly significant increase during early development when larvae were forced to lay on their sides due to their prominent yolk-sacs. In later developmental stages, when they began to swim freely, a dramatic decrease in lapillar asymmetry was apparent. Taken together with own previous findings according to which otolith growth stops after vestibular nerve transaction, the results presented here suggest that the growth and the development of bilateral asymmetry of otoliths is guided by the environmental gravity vector, obviously involving a feedback loop between the brain and the inner ear.

Animals↗

PubMed2000

Electronmicroscopic investigations on the role of vesicle-like bodies in inner ear maculae for fish otolith growth.

The presence, morphology and possible origin of vesicle-like bodies (VBs) within the inner ear otolithic membrane of developmental stages of cichlid fish Oreochromis mossambicus and adult swordtail fish Xiphophorus helleri was analysed by means of transmission and scanning electron microscopy (TEM and SEM, respectively) employing various fixation procedures. The VBs are believed to be involved in the formation of the otolith (or statolith in birds and mammals) regarding the supply of the otolith's organic material. Increasing the osmolarity of the fixation medium decreased the number of VBs seen. Decalcification ended up in a complete disappearance of the VBs. Whilst a fixation with glutaraldehyde followed by OSO4 fixation yielded numerous VBs, only few of them were observed when the tissue was fixed with glutaraldehyde and OSO4 simultaneously. Therefore, the results strongly suggest that the VBs are fixative (i.e., glutaraldehyde) induced artifacts, so-called blisters. With this, the supply of an oto- or statolith's organic material remains obscure. Possibly, it is provided by secretion from the supporting cells as has been hypothesized earlier.

Animals↗

PubMed2000

Gangliosides affect membrane-channel activities dependent on ambient temperature.

1. The functional properties of biological membranes depend on their molecular composition. In regard to this, charged glycosphingolipids play an outstanding role in the functional adaptation of membranes to different temperatures. 2. In order to shed some light on the respective functional properties of complex membraneous glycosphingolipids, the effects of altered temperatures (5-40 degrees C) on planar lipid bilayers made from diphytanoylphosphatidylcholine (DPPC) and alamethicin as an ion channel was analyzed in the presence of either a sialoglycosphingolipid (less polar disialoganglioside GD1a or highly polar tetrasialoganglioside GQ1b) or phosphatidylserine (PS; as control). 3. Different to the control bilayers made from DPPC or DPPC + PS, the bilayers containing gangliosides had specific maxima in alamethicin conductance and stabile life times. Changes in pore-state conductances indicate structural effects based on an interaction of the large (negatively charged) ganglioside headgroups with the alamethicin pores. 4. The results concerning open time and closed time of channels seem to be based on the gangliosides changing the viscosity of the bilayer and possibly introducing phase transitions. 5. Thus, the findings suggest that gangliosides (1) directly affect channel molecules via their headgroups and (2) may additionally affect the fluidity of membranes in order to maintain membrane homeoviscosity in areas surrounding ion channels independent from the environmental temperature. 6. The effects of gangliosides may be of special interest in describing the ability of neuronal adaptation of vertebrates to temperature and more general regarding the functional adaptation of neurons.

Alamethicin↗

PubMed1999

Effect of altered gravity on the neurobiology of fish.

In vertebrates (including humans) altered gravitational environments such as weightlessness can induce malfunction of the inner ears due to a mismatch between canal and statolith afferents. This leads to an illusionary tilt because the inputs from the inner ear are not confirmed by the other sensory organs, which then results in intersensory conflict. Vertebrates in orbit therefore face severe orientation problems. In humans the intersensory conflict may additionally lead to a malaise commonly referred to as space motion sickness (SMS). After the initial days of weightlessness the orientation problems (and SMS) disappear as the brain develops a new interpretation of the available sensory data. The present contribution reviews the neurobiological responses, particularly those of fish, observed under altered gravitational states concerning behavior and neuroplastic reactivities. Investigations employing microgravity (spaceflight, parabolic aircraft flights, clinostat) and hypergravity (laboratory centrifuges as ground-based research tools) provide insights for understanding the basic phenomena, many of which remain only incompletely explained.

Animals↗

PubMed1999

The asymmetrical growth of otoliths in fish is affected by hypergravity.

Size and asymmetry (size difference between the left and the right side) of inner ear otoliths of larval cichlid fish were determined after a long-term stay at moderate hypergravity conditions (3g; centrifuge), in the course of which the animals completed their ontogenetic development from hatch to freely swimming. Both the normal morphogenetic development as well as the timely onset and gain of performance of the swimming behaviour was not impaired by the experimental conditions. However, both utricular and saccular otoliths (lapilli and sagittae, respectively) were significantly smaller after hyper-g exposure as compared to parallely raised 1g control specimens. The asymmetry of sagittae was significantly increased in the experimental animals, whereas the respective asymmetry con-cerning lapilli was pronouncedly decreased in comparison to the 1g controls. These findings suggest, that the growth and the development of bilateral asymmetry of otoliths is guided by the environmental gravity vector.

Animals↗

PubMed1998

Neurobiology of fish under altered gravity conditions.

In vertebrates (including man), an altered gravitational environment such as weightlessness can induce malfunction of the inner ear, based on an irregular dislocation of the otoliths from the corresponding sensory epithelia. This dislocation leads to an illusionary tilt, since the otolithic inputs are not in register with other sensory organs. This results in an intersensory conflict. Vertebrates in orbit therefore face severe orientation problems. In humans, the intersensory conflict may additionally lead to a malaise, commonly referred to as space motion sickness (SMS). During the first days in weightlessness, the orientation problems (and SMS) disappear, since the brain develops a new compensatory interpretation of the available sensory data. The present review reports the neurobiological responses-particularly in fish-observed at altered gravitational states, concerning behaviour and neuroplastic reactivities. Recent investigations employing microgravity (spaceflight, parabolic aircraft flights, clinostat) and hyper-gravity (laboratory centrifuges as ground based research tools) yielded clues and insights into the understanding of the respective basic phenomena. The possible sources of human space sickness (a kinetosis) and of the space adaptation syndrome (when a sensory reinterpretation of gravitational and visual cues takes place) are particularly highlighted with regard to the functional significance of bilaterally asymmetric otoliths (weight, size).

Animals↗

PubMed1998

Neuroplastic reactivity of fish induced by altered gravity conditions: a review of recent results.

A review is being presented concerning behavioural, biochemical, histochemical and electronmicroscopical data on the influence of altered gravitational forces on the swimming performance and on the neuronal differentiation of the brain of cichlid fish larvae and adult swordtail fish that had been exposed to hyper-gravity (3g in laboratory centrifuges), hypo-gravity (>10(-2) g in a fast-rotating clinostat) and to near weightlessness (10(-4) g aboard the Spacelab D-2 mission). After long-term alterations of gravity (and parallel light deprivation), initial disturbances in the swimming behaviour followed by a stepwise regain of normal swimming modes are induced. Parallel, neuroplastic reactivities on different levels of investigation were found, such as adaptive alterations of activities of various enzymes in whole brain as well as in specific neuronal integration centers and an intraneuronal reactivity on ultrastructural level in individual brain parts and in the sensory epithelia of the inner ear. Taken together, these data reveal distinct adaptive neuroplastic reactions of fish to altered gravity conditions.

Adaptation, Physiological↗

PubMed1998

On inappropriately used neuronal circuits as a possible basis of the "loop-swimming" behaviour of fish under reduced gravity: a theoretical study.

One hypothesis for the explanation of the so-called "loop-swimming" behaviour in fish when being subjected to reduced gravity assumes that the activities of the differently weighted otoliths of the two labyrinths are well compensated on ground but that a functional asymmetry is induced in weightlessness, resulting in a tonus asymmetry of the body and by this generating the "loop-swimming" behaviour. The basis of this abnormal behaviour has to be searched for in the central nervous system (cns), where the signal-transduction from the inner ear- related signal internalisation to the signal response takes place. Circuits within the CNS of fish, that could possibly generate the "loop-swimming", might be as follows: An asymmetric activation of vestibulospinal circuits would directly result in a tonus asymmetry of the body. An asymmetric activation of the oculomotor nucleus would generate an asymmetrical rotation of the eyes. This would cause in its turn asymmetric images on the two retinas, which were forwarded to the diencephalic accessory optic system (AOS). It is the task of the AOS to stabilize retinal images, thereby involving the cerebellum, which is the main integration center for sensory and motor modalities. With this, the cerebellar output would generate a tonus asymmetry of the body in order to make the body of the fish follow its eyes. Such movements (especially when assuming an open loop control) would end up in the aforementioned "loop-swimming" behaviour.

Animals↗

PubMed1998

Influence of long-term hyper-gravity on the reactivity of succinic acid dehydrogenase and NADPH-diaphorase in the central nervous system of fish: a histochemical study.

In the course of a densitometric evaluation, the histochemically demonstrated reactivity of succinic acid dehydrogenase (SDH) and of NADPH-diaphorase (NADPHD) was determined in different brain nuclei of two teleost fish (cichlid fish Oreochromis mossambicus, swordtail fish Xiphophorus helleri), which had been kept under 3g hyper-gravity for 8 days. SDH was chosen since it is a rate limiting enzyme of the Krebs cycle and therefore it is regarded as a marker for metabolic and neuronal activity. NADPHD reactivity reflects the activity of nitric oxide synthase. Nitric oxide (NO) is a gaseous intercellular messenger that has been suggested to play a major role in several different in vivo models of neuronal plasticity including learning. Within particular vestibulum-connected brain centers, significant effects of hyper-gravity were obtained, e.g., in the magnocellular nucleus, a primary vestibular relay ganglion of the brain stem octavolateralis area, in the superior rectus subdivision of the oculomotoric nucleus and within cerebellar eurydendroid cells, which in teleosts possibly resemble the deep cerebellar nucleus of higher vertebrates. Non-vestibulum related nuclei did not respond to hyper-gravity in a significant way. The effect of hyper-gravity found was much less distinct in adult animals as compared to the circumstances seen in larval fish (Anken et al., Adv. Space Res. 17, 1996), possibly due to a development correlated loss of neuronal plasticity.

Afferent Pathways↗

PubMed1998

Ultrastructural aspects of otoliths and sensory epithelia of fish inner ear exposed to hypergravity.

The present electron microscopical investigations were directed to the question, whether alterations in the gravitational force might induce structural changes in the morphology of otoliths or/and inner ear sensory epithelia of developing and adult swordtail fish (Xiphophorus helleri) that had been kept either under long-term moderate hypergravity (8 days; 3g) or under short-time extreme hypergravity (10 minutes up to 9g). The otoliths of adult and neonate swordtail fish were investigated by means of scanning electron microscopy (SEM). Macular epithelia of adult fish were examined both by SEM and transmission electron microscopy (TEM). The saccular otoliths (sagittae) of normally hatched adult fish revealed an enormous inter- (and even intra-; i.e. left vs. right) individual diversity in shape and size, whereas the otoliths of utricles (lapilli) and lagenae (asterisci) seemed to be more constant regarding morphological parameters. The structural diversity of juvenile otoliths was found to be less prominent as compared to the adults, differing from the latter regarding their peculiar crystalline morphology. Qualitative differences in the fine structure (SEM) of otoliths taken from adult and larval animals kept under 3g in comparison to 1g controls could not be observed. The SEM and TEM investigations of sensory epithelia also did not reveal any effects due to 3g stimulation. Even extreme hypergravity (more than 7g) for 10 minutes did not result in distinct pathological changes.

Aging↗

PubMed1998

Fish inner ear otolith size and bilateral asymmetry during development.

Size and bilateral asymmetry (i.e. size difference between the left and the right hand side) of inner ear otoliths of larval mouthbreeding cichlid fish were determined during the ontogenetic development of larvae from hatching to the free swimming stage. Animals of two batches were raised in aquarium hatch baskets. The basket containing one batch was placed directly above aeration equipment, resulting in random water circulation within the basket, which constantly shifted the specimens around ('shifted' specimens). The second batch of animals was raised in parallel without shifting. Due to the weight of the yolk-sacs, these animals lay on their sides until the yolk-sacs were resorbed ('stationary' specimens). The groups of larvae did not differ from one another in respect of individual general development, nor in otolith size. Contrasting results were obtained regarding bilateral otolith asymmetry: In both shifted and stationary animals, asymmetry of utricular and saccular otoliths (lapilli and sagittae, respectively) ranged at comparatively low values throughout development. However, by comparison with shifted individuals, lapillar asymmetry of stationary animals showed a highly significant increase during early development when larvae were forced to lay on their sides due to their prominent yolk-sacs. In later developmental stages, when they began to swim freely, a dramatic decrease in lapillar asymmetry was apparent. These findings indicate that development of lapillar asymmetry depends on the direction of the acting gravity vector relative to the positioning of the larvae, suggesting that the size (or mass) of a given otolith is regulated via a feedback mechanism.

Animals↗

PubMed1998

Morphometry of fish inner ear otoliths after development at 3g hypergravity.

Size and asymmetry (size difference between the left and right sides) of inner ear otoliths of larval cichlid fish were determined after a long-term stay in moderate hypergravity conditions (3g; centrifuge), in the course of which the animals completed their ontogenetic development from hatch to freely swimming. Neither the normal morphogenetic development nor the timely onset and gain of performance of swimming behaviour were impaired by the experimental conditions. However, both utricular and saccular otoliths (lapilli and sagittae, respectively) were significantly smaller after hyper-g exposure compared to 1g control specimens raised in parallel. The asymmetry of sagittae was significantly increased in the experimental animals, whereas the respective asymmetry of lapilli was pronouncedly decreased compared with the 1g controls. These findings suggest that growth and development of bilateral asymmetry of otoliths are guided by the environmental gravity vector. Some of the hyper-g animals revealed a kinetotic behaviour on transfer to normal 1g earth conditions, which was similar to the behaviour observed in previous experiments on the transfer from 1g to microgravity (parabolic aircraft flights). The lapillar asymmetry of kinetotic samples was found to be significantly higher than that of normally behaving experimental specimens. No differences in asymmetry of sagittae were obtained between the two groups. This supports an earlier theoretical concept, according to which human static space sickness might be based on asymmetric utricular otoliths.

Animals↗