Ca2+ currents in dorsal unpaired median (DUM) neurons isolated from the fifth abdominal ganglion of the cockroach Periplaneta americana were investigated with the whole cell patch-clamp technique. On the basis of kinetic and pharmacological properties, two different Ca2+ currents were separated in these cells: mid/low-voltage-activated (M-LVA) currents and high-voltage-activated (HVA) currents. M-LVA currents had an activation threshold of -50 mV and reached maximal peak values at -10 mV. They were sensitive to depolarized holding potentials and decayed very rapidly. The decay was largely Ca2+ dependent. M-LVA currents were effectively blocked by Cd2+ median inhibiting concentration (IC50 = 9 microM), but they also had a remarkable sensitivity to Ni2+ (IC50 = 19 microM). M-LVA currents were insensitive to vertebrate LVA channel blockers like flunarizine and amiloride. The currents were, however, potently blocked by omega-conotoxin MVIIC (1 microM) and omega-agatoxin IVA (50 nM). The blocking effects of omega-toxins developed fast (time constant tau = 15 s) and were fully reversible after wash. HVA currents activated positive to -30 mV and showed maximal peak currents at + 10 mV. They were resistant to depolarized holding potentials up to -50 mV and decayed in a less pronounced manner than M-LVA currents. HVA currents were potently blocked by Cd2+ (IC50 = 5 microM) but less affected by Ni2+ (IC50 = 40 microM). These currents were reduced by phenylalkylamines like verapamil (10 microM) and benzothiazepines like diltiazem (10 microM), but they were insensitive to dihydropyridines like nifedipine (10 microM) and BAY K 8644 (10 microM). Furthermore, HVA currents were sensitive to omega-conotoxin GVIA (1 microM). The toxin-induced reduction of currents appeared slowly (tau approximately 120 s) and the recovery after wash was incomplete in most cases. The dihydropyridine insensitivity of the phenylalkylamine-sensitive HVA currents is a property the cockroach DUM cells share with other invertebrate neurons. Compared with Ca2+ currents in vertebrates, the DUM neuron current differ considerably from the presently known types. Although there are some similarities concerning kinetics, the pharmacological profile of the cockroach Ca2+ currents especially is very different from profiles already described for vertebrate currents.
An antiserum was raised in rabbits immunized with octopamine conjugated to thyroglobulin. The specificity of this antiserum for octopamine is shown by dot blot immunoassay analysis. The antiserum does not crossreact with dopamine, noradrenaline, and serotonin, but slight crossreactivity with the amine tyramine at high concentrations was observed. The tyramine crossreactivity could be eliminated by preabsorption with a tyramine-glutaraldehyde-BSA conjugate. Using this antiserum, we describe the topographical distribution of octopamine-immunoreactive (ir) neuronal elements in wholemounts and paraffin sections of the ventral nerve cord of the American cockroach. The pattern of octopamine immunostaining is completely different from that obtained with an antidopamine serum, and can be blocked by preabsorbing the antioctopamine serum with BSA-conjugated octopamine. Cell bodies and dendritic processes of putatively octopaminergic dorsal (DUM) and ventral (VUM) unpaired median neurons were clearly octopamine-ir in all ganglia examined. The numbers of stained DUM somata in the mesothoracic, metathoracic, and terminal ganglion of females correspond to those of peripherally projecting DUM cells revealed previously by retrograde tracing (Gregory, Philos Trans R Soc Lond [Biol] 306:191, 1984; Tanaka and Washio, Comp Biochem Physiol 91A:37, 1988; Stoya et al., Zool Jb Physiol 93:75, 1989). In addition, various, previously unknown, paired cells with octopamine-like immunoreactivity were found in all ventral ganglia except abdominal ganglia 3-6. Some of these probably project intersegmentally.
The octapeptide neurohormone D (NHD), a member of the family of adipokinetic hormones (AKH-peptides), increases the frequency of spontaneous activity in dorsal unpaired median (DUM) neurones isolated from the terminal ganglion of the cockroach Periplaneta americana. The increase in spike frequency is accompanied by changes in the shape and the amplitude of the single action potentials, e.g. a more pronounced afterhyperpolarization. Effects of NHD on membrane currents were investigated in these DUM cells with whole-cell voltage-clamp measurements. A voltage-independent Ca2+ current flowing at the resting potential (ICa,R) was found. NHD, at nanomolar concentrations, enhanced this ICa,R in a concentration-dependent manner. 0.1 mM Cd2+ markedly reduced ICa,R and in this case ICa,R was hardly potentiated by NHD. In the presence of NHD a fast activating Ca(2+)-dependent K+ current sensitive to charybdotoxin and to low concentrations of tetraethylammonium was augmented. The enhanced afterhyperpolarization of action potentials can be accounted for by the increase in the Ca(2+)-dependent K+ current. The changes of the membrane currents induced by NHD are discussed with respect to further effects on the spike pattern and in relation to the previously described mode of action of AKH-peptides in other preparations.
To approach the question of what life is, we first have to state that life exists exclusively as the "being-alive" of discrete spatio-temporal entities. The simplest "unit" that can legitimately be considered to be alive is an intact prokaryotic cell as a whole. In this review, I discuss critically various aspects of the nature and singularity of living beings from the biologist's point of view. In spite of the enormous richness of forms and performances in the biotic realm, there is a considerable uniformity in the chemical "machinery of life," which powers all organisms. Life represents a dynamic state; it is performance of a system of singular kind: "life-as-action" approach. All "life-as-things" hypotheses are wrong from the beginning. Life is conditioned by certain substances but not defined by them. Living systems are endowed with a power to maintain their inherent functional order (organization) permanently against disruptive influences. The term organization inherently involves the aspect of functionality, the teleonomic, purposeful cooperation of structural and functional elements. Structures in turn require information for their specification, and information presupposes a source. This source is constituted in living systems by the nucleic acids. Organisms are unique in having a capacity to use, maintain, and replicate internal information, which yields the basis for their specific organization in its perpetuation. The existence of a genome is a necessary condition for life and one of the absolute differences between living and non-living matter. Organization includes both what makes life possible and what is determined by it. It is not something "implanted" into the living beings but has its origin and capacity for maintenance within the system itself. It is the essence of life. The property of being alive we can consider as an emergent property of cells that corresponds to a certain level of self-maintained complex order or organization.
In the American cockroach, the distribution and connections of neuronal elements of the terminal ganglion-proctodeal nerve-hindgut system were investigated by means of immunohistochemical methods and axonal CoCl2 iontophoresis. Proctolinlike immunoreactivity was localized within neurons of the terminal ganglion projecting into the proctodeal nerve on the one hand, and in nerve cells without a direct connection to this system on the other. Immunohistochemically, in whole mount preparations fibres of the proctodeal nerve and terminal structures in the hindgut musculature exhibit strong proctolinlike immunoreactivity. At the light- and electron-microscopic levels the pathways of about 30 somata of the proctodeal neural system were characterized by cobalt chloride iontophoresis. The relationships of cobalt filled and immunoreactive neuronal structures are discussed.
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