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1.
Dokl Biol Sci ; 505(1): 105-108, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-36038796

RESUMO

A total of 1250 lower first molars (m1) of voles (Arvicolini) were studied from Late Pleistocene deposits (the radiocarbon dates from rodent bones: 17 100 ± 50 IGANAMS-9117; 13 255 ± 60 IGANAMS-9116) of the Imanay Cave (southern Urals, 53°02' N, 56°26' E). Of these, 24 m1 of voles of the subgenus Stenocranius were found to have broadly connected triangles T4 and T5 (Pitymys-Rhombus) at the base of the anteroconid. This structure is characteristic of lower m1 of Lasiopodomys (Stenocranius) gregaloides (Hinton, 1923) from faunas of the second half of the Early Pleistocene and the first half of the Middle Pleistocene. Molars with such morphology have not been found so far in the Late Pleistocene and Holocene locations of Europe and the Urals.


Assuntos
Arvicolinae , Roedores , Animais , Cavernas , Europa (Continente) , Fósseis
2.
Dokl Biol Sci ; 481(1): 160-162, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30171471

RESUMO

The study deals with a new sample of the mammalian fossils from the Pleistocene deposits of the Ignatievskaya Cave in Southern Urals (54°53' N, 57°46' E). Among the rodent fossils, the teeth of red squirrel (Sciurus vulgaris) and forest dormouse (Dryomys nitedula) have been identified. The enamel differentiation quotient (SDQ) of the water vole teeth matches the Arvicola terrestris species from locations of the end of Middle and Late Pleistocene (oxygen isotope stages, OIS 6 and OIS 5-2, respectively). The high SDQ value for the water vole and the presence of the squirrel and dormouse in the fauna allow dating the finding near the end of the Middle Pleistocene and beginning of the Late Pleistocene (OIS 6 and OIS 5e, respectively). The widespread open and forest landscapes were inhabited by this fauna.


Assuntos
Fósseis , Roedores , Sciuridae , Animais
3.
Dokl Biol Sci ; 482(1): 191-193, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30402757

RESUMO

The vertebrate fauna from the cave deposits in Imanai Cave in the Southern Urals (53°02' N, 56°26'E) has been studied. It contains 715 bones that belonged to at least 11 individuals of fossil lion (Panthera (Leo) ex gr. fossilis-spelaea). It has been established that this is one of the largest Eurasian burial sites of fossil lions. The bones were accumulated due to the natural death of animals inside the cave. The age and sex estimations have shown that at least six adult males and five adult females died there. According to the accompanying fauna, radiocarbon, geochemical, and mineralogical analyses and archaeological finds, the interval of the lion bone accumulation is determined as the first half to middle of Late Pleistocene (OIS 5-3).


Assuntos
Fósseis/anatomia & histologia , Leões/anatomia & histologia , Animais , Osso e Ossos/anatomia & histologia , Cavernas , Feminino , Masculino
4.
Biochemistry (Mosc) ; 82(8): 906-915, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28941458

RESUMO

Energy relaxation was studied with difference femtosecond spectroscopy in reaction centers of the YM210L mutant of the purple photosynthetic bacterium Rhodobacter sphaeroides at low temperature (90 K). A dynamical long-wavelength shift of stimulated emission of the excited state of the bacteriochlorophyll dimer P was found, which starts simultaneously with P* formation and is accompanied by a change in the spectral shape of this emission. The characteristic value of this shift was about 30 nm, and the characteristic time about 200 fs. Difference kinetics ΔA measured at fixed wavelengths demonstrate the femtosecond shift of the P* stimulated emission appearing as a dependence of these kinetics on wavelength. We found that the reported long-wavelength shift can be explained in terms of electron-vibrational relaxation of the P* excited state with time constants of vibrational and electronic relaxation of 100 and 50 fs, respectively. Alternative mechanisms of the dynamical shift of the P* stimulated emission spectrum are also discussed in terms of energy redistribution between vibrational modes or coherent excitation of the modes.


Assuntos
Proteínas de Bactérias/metabolismo , Complexo de Proteínas do Centro de Reação Fotossintética/metabolismo , Rhodobacter sphaeroides/metabolismo , Proteínas de Bactérias/genética , Bacterioclorofilas/química , Bacterioclorofilas/metabolismo , Dimerização , Cinética , Lasers de Estado Sólido , Mutagênese Sítio-Dirigida , Complexo de Proteínas do Centro de Reação Fotossintética/genética , Rhodobacter sphaeroides/efeitos da radiação , Espectrofotometria
5.
Artigo em Russo | MEDLINE | ID: mdl-26259278

RESUMO

There have been presented analysis of ornithosis outbreaks among population of Orenburg and Kurgan regions in 2008-2009. Find out factors and conditions that promote conducive of epidemic foci. Have been presented a list of the main preventive measures.


Assuntos
Reservatórios de Doenças , Psitacose/epidemiologia , Zoonoses/epidemiologia , Animais , Aves/microbiologia , Aves/virologia , Surtos de Doenças , Humanos , Psitacose/patologia , Federação Russa , Zoonoses/patologia
6.
J Theor Biol ; 343: 92-101, 2014 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-24270095

RESUMO

Primary charge separation in reaction centers (RCs) of bacterial photosynthesis is modeled in this work. An incoherent population dynamics of RCs states is formulated by kinetic equations. It is assumed that charge separation is accompanied by regular motion of the system along additional coordinates. This motion modulates an energetics of the reactions, and this modulation causes femtosecond oscillations in the population of the states. The best qualitative and quantitative accordance with experimental data on native, modified and mutant RCs of Rba. sphaeroides is achieved in the five states model that includes two excited states P(*)905BAHA and P(*)940BAHA and three charge separated states I, P(+)BA(-)HA and P(+)BAHA(-) (P is a primary electron donor, bacteriochlorophyll dimer, BA and HA are electron acceptors, monomeric bacteriochlorophyll and bacteriopheophytin in active A-branch respectively). The excited states emit at 905 and 940 nm and have approximately the same energy and high interaction rate. The intermediate state I is populated earlier than the P(+)BA(-)HA state and has energy close to the energy of the excited states, a high rate of population and depopulation and spectral identity to the BA(-). A sum of the I and P(+)BA(-)HA populations fits the experimental kinetics of the BA(-) absorption band at 1020 nm. The model explains an oscillatory phenomenon in the kinetics of the P(*) stimulated emission and of the BA(-) absorption. In the schemes without the I state, accordance with the experiment is achieved at unreal parameter values or is not achieved at all. A qualitative agreement of the model with the experiment can be achieved at a wide range of parameter values. The nature of the states I and P(*)940BAHA is discussed in terms of partial charge separation between P and BA and inside P respectively.


Assuntos
Elétrons , Modelos Biológicos , Complexo de Proteínas do Centro de Reação Fotossintética/metabolismo , Cinética , Rhodobacter sphaeroides/metabolismo , Termodinâmica
7.
Biochemistry (Mosc) ; 79(3): 251-9, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24821452

RESUMO

This work continuous a series of studies devoted to discovering principles of organization of natural antennas in photosynthetic microorganisms that generate in vivo large and highly effective light-harvesting structures. The largest antenna is observed in green photosynthesizing bacteria, which are able to grow over a wide range of light intensities and adapt to low intensities by increasing of size of peripheral BChl c/d/e antenna. However, increasing antenna size must inevitably cause structural changes needed to maintain high efficiency of its functioning. Our model calculations have demonstrated that aggregation of the light-harvesting antenna pigments represents one of the universal structural factors that optimize functioning of any antenna and manage antenna efficiency. If the degree of aggregation of antenna pigments is a variable parameter, then efficiency of the antenna increases with increasing size of a single aggregate of the antenna. This means that change in degree of pigment aggregation controlled by light-harvesting antenna size is biologically expedient. We showed in our previous work on the oligomeric chlorosomal BChl c superantenna of green bacteria of the Chloroflexaceae family that this principle of optimization of variable antenna structure, whose size is controlled by light intensity during growth of bacteria, is actually realized in vivo. Studies of this phenomenon are continued in the present work, expanding the number of studied biological materials and investigating optical linear and nonlinear spectra of chlorosomes having different structures. We show for oligomeric chlorosomal superantennas of green bacteria (from two different families, Chloroflexaceae and Oscillochloridaceae) that a single BChl c aggregate is of small size, and the degree of BChl c aggregation is a variable parameter, which is controlled by the size of the entire BChl c superantenna, and the latter, in turn, is controlled by light intensity in the course of cell culture growth.


Assuntos
Proteínas de Bactérias/química , Complexos de Proteínas Captadores de Luz/química , Luz , Proteínas de Bactérias/metabolismo , Bacterioclorofilas/química , Chloroflexi/metabolismo , Chloroflexus/metabolismo , Complexos de Proteínas Captadores de Luz/metabolismo , Fotossíntese/fisiologia
8.
Biochemistry (Mosc) ; 77(5): 444-55, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22813585

RESUMO

In this work, the influence of the crystallographic water on electron transfer between primary donor P and acceptor B(A) was studied in reaction centers (RCs) of the purple bacterium Rhodobacter sphaeroides and the green bacterium Chloroflexus aurantiacus. For this purpose, time constants and oscillations of charge separation kinetics are compared between dry film RCs and RCs in glycerol-water buffer at 90 K. A common result of the drying of Rba. sphaeroides and Cfx. aurantiacus RCs is slowing of the charge separation process, decrease in amplitude of the oscillatory components of the kinetics, and the depletion of its spectrum. Thus, the major time constant of stimulated emission decay of P* bacteriochlorophyll dimer at 940 nm is increased from 1.1 psec for water-containing Rba. sphaeroides RCs to 1.9 psec for dry films of Rba. sphaeroides RCs. An analogous increase from 3.5 to 4.2 psec takes place in Cfx. aurantiacus RCs. In dry films of Rba. sphaeroides RCs, the amplitude of coherent oscillations of the absorption band of monomeric bacteriochlorophyll B(A)(-) at 1020 nm is 1.8 times less for the 130-cm(-1) component and 2.3 times less for the 32-cm(-1) component than the analogous amplitudes for water-containing RCs. Measurements in the analogous band of Cfx. aurantiacus RCs show that strong decrease (~5-10 times) of the B(A)(-) absorption band and strong slowing (from ~0.8 to ~3 psec) of B(A)(-) accumulation together with ~3-fold decrease in oscillation amplitude occurs on drying of these RCs. The overtones of the 32-cm(-1) component disappeared from the oscillations of the kinetics at 940 and 1020-1028 nm after drying of the Rba. sphaeroides and Cfx. aurantiacus RCs. The results are in agreement with the results for GM203L mutant of Rba. sphaeroides, in which the HOH55 water molecule is sterically removed, and with the results for dry films of pheophytin-modified RCs of Rba. sphaeroides R-26 and for YM210W and YM210L Rba. sphaeroides mutant RCs. The data are discussed in terms of the influence (or participation) of the HOH55 water molecule on electron transfer along the chain of polar atomic groups N-Mg(P(B))-N-C-N(HisM202)-HOH55-O=(B(A)) connecting P(B) and B(A) in Rba. sphaeroides RCs.


Assuntos
Chloroflexus/metabolismo , Complexo de Proteínas do Centro de Reação Fotossintética/metabolismo , Rhodobacter sphaeroides/metabolismo , Transporte de Elétrons , Cinética , Complexo de Proteínas do Centro de Reação Fotossintética/química , Estrutura Terciária de Proteína , Espectroscopia de Infravermelho com Transformada de Fourier , Temperatura , Fatores de Tempo
9.
Biochemistry (Mosc) ; 76(10): 1107-19, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22098236

RESUMO

Primary stage of charge separation and transfer of charges was studied in reaction centers (RCs) of point mutants LL131H and LL131H/LM160H/FM197H of the purple bacterium Rhodobacter sphaeroides by differential absorption spectroscopy with temporal resolution of 18 fsec at 90 K. Difference absorption spectra measured at 0-4 psec delays after excitation of dimer P at 870 nm with 30 fsec step were obtained in the spectral range of 935-1060 nm. It was found that a decay of P* due to charge separation is considerably slower in the mutant RCs in comparison with native RCs of Rba. sphaeroides. Coherent oscillations were found in the kinetics of stimulated emission of the P* state at 940 nm. Fourier analysis of the oscillations revealed a set of characteristic bands in the frequency range of 20-500 cm(-1). The most intense band has the frequency of ~130 cm(-1) in RCs of mutant LL131H and in native RCs and the frequency of ~100 cm(-1) in RCs of the triple mutant. It was found that an absorption band of bacteriochlorophyll anion B(A)(-) which is registered in the difference absorption spectra of native RCs at 1020 nm is absent in the analogous spectra of the mutants. The results are analyzed in terms of the participation of the B(A) molecule in the primary electron transfer in the presence of a nuclear wave packet moving along the inharmonic surface of P* potential energy.


Assuntos
Fotossíntese , Complexo de Proteínas do Centro de Reação Fotossintética/química , Rhodobacter sphaeroides/enzimologia , Bacterioclorofilas/química , Bacterioclorofilas/metabolismo , Transporte de Elétrons , Cinética , Mutação , Complexo de Proteínas do Centro de Reação Fotossintética/genética , Complexo de Proteínas do Centro de Reação Fotossintética/metabolismo , Espectroscopia por Absorção de Raios X
10.
Biochemistry (Mosc) ; 75(4): 412-22, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-20618129

RESUMO

Coherent processes in an initial phase of charge transfer in reaction centers (RCs) of the triple mutant S(L178)K/G(M203)D/L(M214)H of Rhodobacter sphaeroides were investigated by difference (light - dark) absorption spectroscopy with 18 fsec time resolution. Electron transfer in the B cofactor branch is activated in this mutant, while the A-branch electron transfer is slowed in comparison with native RCs of Rba. sphaeroides. A bulk of absorption difference spectra was analyzed in the 940-1060 nm range (stimulated emission of excited bacteriochlorophyll dimer P* and absorption of bacteriochlorophyll anions B(A)(-) and beta(-), where beta is a bacteriochlorophyll substituting the native bacteriopheophytin H(A)) and in the 735-775 nm range (bleaching of the absorption band of the bacteriopheophytin H(B) in the B-branch) in the -0.1 to 4 psec range of delays with respect to the moment of photoexcitation of P at 870 nm. Spectra were measured at 293 and 90 K. The kinetics of P* stimulated emission at 940 nm shows its decay with a time constant of approximately 14 psec at 90 K and approximately 18 psec at 293 K, which is accompanied by oscillations with a frequency of approximately 150 cm(-1). A weak absorption band is found at 1018 nm that is formed approximately 100 fsec after excitation of P and reflects the electron transfer from P* to beta and/or B(A) with accumulation of the P(+)beta(-) and/or P(+)B(A)(-) states. The kinetics of DeltaA at 1018 nm contains the oscillations at approximately 150 cm(-1) and distinct low-frequency oscillations at 20-100 cm(-1); also, the amplitude of the oscillations at 150 cm(-1) is much smaller at 293 than at 90 K. The oscillations in the kinetics of the 1018 nm band do not contain a 32 cm(-1) mode that is characteristic for native Rba. sphaeroides RCs having water molecule HOH55 in their structure. The DeltaA kinetics at 751 nm reflects the electron transfer to H(B) with formation of the P(+)H(B)(-) state. The oscillatory part of this kinetics has the form of a single peak with a maximum at ~50 fsec completely decaying at ~200 fsec, which might reflect a reversible electron transfer to the B-branch. The results are analyzed in terms of coherent nuclear wave packet motion induced in the P* excited state by femtosecond light pulses, of an influence of the incorporated mutations on the mutual position of the energy levels of charge separated states, and of the role of water HOH55 in the dynamics of the initial electron transfer.


Assuntos
Proteínas de Bactérias/química , Complexo de Proteínas do Centro de Reação Fotossintética/química , Rhodobacter sphaeroides/enzimologia , Substituição de Aminoácidos , Proteínas de Bactérias/genética , Bacterioclorofilas/química , Transporte de Elétrons , Cinética , Mutação , Complexo de Proteínas do Centro de Reação Fotossintética/genética , Fatores de Tempo
11.
Biochemistry (Mosc) ; 75(7): 832-40, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20673206

RESUMO

The role of tyrosine M210 in charge separation and stabilization of separated charges was studied by analyzing of the femtosecond oscillations in the kinetics of decay of stimulated emission from P* and of a population of the primary charge separated state P(+)B(A)(-) in YM210L and YM210L/HL168L mutant reaction centers (RCs) of Rhodobacter sphaeroides in comparison with those in native Rba. sphaeroides RCs. In the mutant RCs, TyrM210 was replaced by Leu. The HL168L mutation placed the redox potential of the P(+)/P pair 123 mV below that of native RCs, thus creating a theoretical possibility of P(+)B(A)(-) stabilization. Kinetics of P* decay at 940 nm of both mutants show a significant slowing of the primary charge separation reaction in comparison with native RCs. Distinct damped oscillations in these kinetics with main frequency bands in the range of 90-150 cm(-1) reflect mostly nuclear motions inside the dimer P. Formation of a very small absorption band of B(A)(-) at 1020 nm is registered in RCs of both mutants. The formation of the B(A)(-) band is accompanied by damped oscillations with main frequencies from ~10 to ~150 cm(-1). Only a partial stabilization of the P(+)B(A)(-) state is seen in the YM210L/HL168L mutant in the form of a small non-oscillating background of the 1020-nm kinetics. A similar charge stabilization is absent in the YM210L mutant. A model of oscillatory reorientation of the OH-group of TyrM210 in the electric fields of P(+) and B(A)(-) is proposed to explain rapid stabilization of the P(+)B(A)(-) state in native RCs. Small oscillatory components at ~330-380 cm(-1) in the 1020-nm kinetics of native RCs are assumed to reflect this reorientation. We conclude that the absence of TyrM210 probably cannot be compensated by lowering of the P(+)B(A)(-) free energy that is expected for the double YM210L/HL168L mutant. An oscillatory motion of the HOH55 water molecule under the influence of P(+) and B(A)(-) is assumed to be another potential contributor to the mechanism of P(+)B(A)(-) stabilization.


Assuntos
Proteínas de Bactérias/metabolismo , Mutação de Sentido Incorreto , Complexo de Proteínas do Centro de Reação Fotossintética/metabolismo , Rhodobacter sphaeroides/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Transporte de Elétrons , Cinética , Oxirredução , Complexo de Proteínas do Centro de Reação Fotossintética/química , Complexo de Proteínas do Centro de Reação Fotossintética/genética , Rhodobacter sphaeroides/química , Rhodobacter sphaeroides/genética
13.
Biochemistry (Mosc) ; 74(11): 1203-10, 2009 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-19916934

RESUMO

Difference femtosecond absorption spectroscopy with 20-fsec temporal resolution was applied to study a primary stage of charge separation and transfer processes in reaction centers of YM210L and YM210L/FM197Y site-directed mutants of the purple bacterium Rhodobacter sphaeroides at 90 K. Photoexcitation was tuned to the absorption band of the primary electron donor P at 880 nm. Coherent oscillations in the kinetics of stimulated emission of P* excited state at 940 nm and of anion absorption of monomeric bacteriochlorophyll B(A)(-) at 1020 nm were monitored. The absence of tyrosine YM210 in RCs of both mutants leads to strong slowing of the primary reaction P* --> P(+)B(A)(-) and to the absence of stabilization of separated charges in the state P(+)B(A)(-). Mutation FM197Y increases effective mass of an acetyl group of pyrrole ring I in the bacteriochlorophyll molecule P(B) of the double mutant YM210L/FM197Y by a hydrogen bond with OH-TyrM197 group that leads to a decrease in the frequency of coherent nuclear motions from 150 cm(-1) in the single mutant YM210L to ~100 cm(-1) in the double mutant. Oscillations with 100-150 cm(-1) frequencies in the dynamics of the P* stimulated emission and in the kinetics of the reversible formation of P(+)B(A)(-) state of both mutants reflect a motion of the P(B) molecule relatively to P(A) in the area of mutual overlapping of their pyrrole rings I. In the double mutant YM210L/FM197Y the oscillations in the P* emission band and the B(A)(-) absorption band are conserved within a shorter time ~0.5 psec (1.5 psec in the YM210L mutant), which may be a consequence of an increase in the number of nuclei forming a wave packet by adding a supplementary mass to the dimer P.


Assuntos
Mutação , Complexo de Proteínas do Centro de Reação Fotossintética/química , Rhodobacter sphaeroides/química , Rhodobacter sphaeroides/genética , Bacterioclorofilas/química , Cinética , Complexo de Proteínas do Centro de Reação Fotossintética/genética , Complexo de Proteínas do Centro de Reação Fotossintética/metabolismo , Rhodobacter sphaeroides/metabolismo , Análise Espectral
14.
Biochemistry (Mosc) ; 74(8): 846-54, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19817684

RESUMO

Difference absorption spectroscopy with temporal resolution of approximately 20 fsec was used to study the primary phase of charge separation in isolated reaction centers (RCs) of Chloroflexus aurantiacus at 90 K. An ensemble of difference (light-minus-dark) absorption spectra in the 730-795 nm region measured at -0.1 to 4 psec delays relative to the excitation pulse was analyzed. Comparison with analogous data for RCs of HM182L mutant of Rhodobacter sphaeroides having the same pigment composition identified the 785 nm absorption band as the band of bacteriopheophytin Phi(B) in the B-branch. By study the bleaching of this absorption band due to formation of Phi(B)(-), it was found that a coherent electron transfer from P* to the B-branch occurs with a very small delay of 10-20 fsec after excitation of dimer bacteriochlorophyll P. Only at 120 fsec delay electron transfer from P* to the A-branch occurs with the formation of bacteriochlorophyll anion B(A)(-) absorption band at 1028 nm and the appearance of P* stimulated emission at 940 nm, as also occurs in native RCs of Rb. sphaeroides. It is concluded that a nuclear wave packet motion on the potential energy surface of P* after a 20-fsec light pulse excitation leads to the coherent formation of the P(+)Phi(B)(-) and P(+)B(A)(-) states.


Assuntos
Chloroflexus/química , Complexo de Proteínas do Centro de Reação Fotossintética/química , Chloroflexus/metabolismo , Transporte de Elétrons , Cinética , Complexo de Proteínas do Centro de Reação Fotossintética/metabolismo , Análise Espectral
15.
Biophys Chem ; 240: 1-8, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29857169

RESUMO

In green photosynthetic bacteria, the chlorosomal bacteriochlorophyll molecules are organized via self-assembly and do not require proteins to provide a scaffold for efficient light harvesting. Despite numerous investigations, a consensus regarding the spatial structure of chlorosomal antennae has not yet been reached. For the first time, we demonstrated by coherent femtosecond spectroscopy at cryogenic temperature that the very low-frequency (~101 cm-1) vibrations of bacteriochlorophyll c pigments in isolated Chloroflexus aurantiacus chlorosomes are sensitive to their oligomerisation extent which depends on the light intensity during the growth of the cell cultures. We explained this sensitivity in terms of the coupling of delocalised vibration modes of BChl c molecules aggregated into chains within their antenna unit building blocks. These findings, together with previously obtained spectroscopy and microscopy data, confirmed that the unit building blocks functioning within Chloroflexus aurantiacus chlorosomal antenna are built up from the rather short (2-5 BChl c pigments) quasi-linear chains. The approach presented here seems to be perspective since it directly reveals structural and dynamical properties of the oligomeric systems.


Assuntos
Proteínas de Bactérias/metabolismo , Bacterioclorofilas/metabolismo , Chloroflexus/metabolismo , Proteínas de Bactérias/química , Bacterioclorofilas/química , Cinética , Luz , Polimerização , Temperatura , Vibração
16.
J Neurosci ; 21(19): 7439-46, 2001 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-11567033

RESUMO

Neuronal apoptosis plays an essential role in early brain development and contributes to secondary neuronal loss after acute brain injury. Recent studies have provided evidence that neuronal susceptibility to apoptosis induced by traumatic or ischemic injury decreases during brain development. However, the molecular mechanisms responsible for this age-dependent phenomenon remain unclear. Here we demonstrate that, during brain maturation, the potential of the intrinsic apoptotic pathway is progressively reduced and that such repression is associated with downregulation of apoptotic protease-activating factor-1 (Apaf-1) and caspase-3 gene expression. A similar decline in apoptotic susceptibility associated with downregulation of Apaf-1 expression as a function of developmental age was also found in cultured primary rat cortical neurons. Injury-induced cytochrome c-specific cleavage of caspase-9 followed by activation of caspase-3 in mature brain correlated with marked increases in Apaf-1 and caspase-3 mRNA and protein expression. These results suggest that differential expression of Apaf-1 and caspase-3 genes may underlie regulation of apoptotic susceptibility during brain development, as well as after acute injury to mature brain, through the intrinsic pathway of caspase activation.


Assuntos
Apoptose , Lesões Encefálicas/metabolismo , Encéfalo/metabolismo , Caspases/biossíntese , Regulação da Expressão Gênica no Desenvolvimento , Biossíntese de Proteínas , Envelhecimento/metabolismo , Animais , Fator Apoptótico 1 Ativador de Proteases , Western Blotting , Encéfalo/embriologia , Encéfalo/crescimento & desenvolvimento , Lesões Encefálicas/patologia , Caspase 3 , Caspases/genética , Sobrevivência Celular/fisiologia , Sistema Livre de Células , Células Cultivadas , Córtex Cerebral/química , Córtex Cerebral/citologia , Córtex Cerebral/metabolismo , Grupo dos Citocromos c/metabolismo , Citosol/metabolismo , Nucleotídeos de Desoxiadenina/metabolismo , Modelos Animais de Doenças , Ativação Enzimática/fisiologia , Masculino , Proteínas/genética , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/fisiologia , Ferimentos não Penetrantes
17.
Cell Death Differ ; 11(10): 1121-32, 2004 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-15375383

RESUMO

Anandamide (arachidonoylethanolamide or AEA) is an endocannabinoid that acts at vanilloid (VR1) as well as at cannabinoid (CB1/CB2) and NMDA receptors. Here, we show that AEA, in a dose-dependent manner, causes cell death in cultured rat cortical neurons and cerebellar granule cells. Inhibition of CB1, CB2, VR1 or NMDA receptors by selective antagonists did not reduce AEA neurotoxicity. Anandamide-induced neuronal cell loss was associated with increased intracellular Ca(2+), nuclear condensation and fragmentation, decreases in mitochondrial membrane potential, translocation of cytochrome c, and upregulation of caspase-3-like activity. However, caspase-3, caspase-8 or caspase-9 inhibitors, or blockade of protein synthesis by cycloheximide did not alter anandamide-related cell death. Moreover, AEA caused cell death in caspase-3-deficient MCF-7 cell line and showed similar cytotoxic effects in caspase-9 dominant-negative, caspase-8 dominant-negative or mock-transfected SH-SY5Y neuroblastoma cells. Anandamide upregulated calpain activity in cortical neurons, as revealed by alpha-spectrin cleavage, which was attenuated by the calpain inhibitor calpastatin. Calpain inhibition significantly limited anandamide-induced neuronal loss and associated cytochrome c release. These data indicate that AEA neurotoxicity appears not to be mediated by CB1, CB2, VR1 or NMDA receptors and suggest that calpain activation, rather than intrinsic or extrinsic caspase pathways, may play a critical role in anandamide-induced cell death.


Assuntos
Apoptose/efeitos dos fármacos , Ácidos Araquidônicos/farmacologia , Calpaína/metabolismo , Caspases/metabolismo , Neurônios/citologia , Neurônios/efeitos dos fármacos , Amidoidrolases/antagonistas & inibidores , Amidoidrolases/metabolismo , Animais , Ácidos Araquidônicos/química , Ácidos Araquidônicos/metabolismo , Cálcio/metabolismo , Calpaína/antagonistas & inibidores , Antagonistas de Receptores de Canabinoides , Inibidores de Caspase , Caspases/deficiência , Caspases/genética , Células Cultivadas , Citocromos c/metabolismo , Endocanabinoides , Ativação Enzimática , Inibidores Enzimáticos/farmacologia , Humanos , Potenciais da Membrana/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Neurônios/metabolismo , Alcamidas Poli-Insaturadas , Transporte Proteico , Ratos , Receptores de Canabinoides/metabolismo , Receptores de Droga/antagonistas & inibidores , Receptores de Droga/metabolismo , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Receptores de N-Metil-D-Aspartato/metabolismo , Transdução de Sinais
18.
Mol Neurobiol ; 24(1-3): 131-44, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-11831549

RESUMO

Recent studies have suggested a role for neuronal apoptosis in cell loss following acute CNS injury as well as in chronic neurodegeneration. Caspases are a family of cysteine requiring aspartate proteases with sequence similarity to Ced-3 protein of Caenorhabditis elegans. These proteases have been found to contribute significantly to the morphological and biochemical manifestations of apoptotic cell death. Caspases are translated as inactive zymogens and become active after specific cleavage. Of the 14 identified caspases, caspase-3 appears to be the major effector of neuronal apoptosis induced by a variety of stimuli. A role for caspase-3 in injury-induced neuronal cell death has been established using semispecific peptide caspase inhibitors. This article reviews the current literature relating to pathways regulating caspase activation in apoptosis associated with acute and chronic neurodegeneration, and suggests that identification of critical upstream caspase regulatory mechanisms may permit more effective treatment of such disorders.


Assuntos
Apoptose/fisiologia , Lesões Encefálicas/patologia , Caspases/metabolismo , Traumatismos da Medula Espinal/patologia , Animais , Ativação Enzimática , Humanos
19.
Gene ; 155(2): 201-5, 1995 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-7721091

RESUMO

The third intron of the mitochondrial benzodiazepine receptor (MBR)-encoding gene was sequenced from hamster, mouse and human. The rodent species were found to include an Alu-like sequence, as was first discovered in the rat gene. Differences with the rat intron were evident by an insertion of an additional B1 element in the hamster and the introduction of a complete and two partial B2 sequences in the mouse intron. The human intron contained a cluster of four Alu sequences; however, all of these repetitive elements were found to be in the opposite orientation relative to the Alu-like sequence present in the rodent genes. These findings support the possibility that the rodent Alu-like sequence is a remnant of a retropositional insertion in this gene prior to the divergence of rodent species. Because the human intron does not contain the same Alu remnant, it cannot be concluded that the rodent sequence represents an insertion of a primordial Alu element prior to the divergence of rodent and primate lineages.


Assuntos
Receptores de GABA-A/genética , Sequências Repetitivas de Ácido Nucleico/genética , Animais , Sequência de Bases , Cricetinae , Cães , Genoma , Humanos , Íntrons/genética , Mitocôndrias/metabolismo , Dados de Sequência Molecular , Ratos , Alinhamento de Sequência , Análise de Sequência
20.
Gene ; 121(2): 377-82, 1992 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-1332914

RESUMO

The gene encoding the rat mitochondrial benzodiazepine receptor (MBR) was cloned and characterized. Hybridization of a previously cloned cDNA for MBR to genomic Southern blots indicated that the gene was probably present at one copy per haploid genome. Rapid amplification of cDNA ends with rat adrenal RNA was used to obtain 47 nt of additional sequence upstream from our previously cloned MBR cDNA proving to be a crucial step in cloning the first exon of this gene. The MBR gene is comprised of four exons spanning approx. 10 kb. The first intron, contained within a 8-kb stretch of this gene, is located within the 5'-untranslated sequence, whereas the remaining two introns are much shorter (641 and 854 bp) and interrupt the coding sequence. The third intron contains sequences homologous to rodent B1 repetitive elements and a novel sequence closely resembling part of a repetitive element belonging to the Alu family in humans. The transcription start point was mapped by S1 nuclease protection assays suggesting that the first exon is just 56 bp in length. The sequence upstream from this region contains three GC boxes but lacks other known consensus recognition sites for sequence-specific transcription factors.


Assuntos
DNA Mitocondrial/genética , Receptores de GABA-A/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Genes , Íntrons , Mitocôndrias/química , Dados de Sequência Molecular , RNA Mensageiro/genética , Ratos , Sequências Reguladoras de Ácido Nucleico , Sequências Repetitivas de Ácido Nucleico , Mapeamento por Restrição , Alinhamento de Sequência , Transcrição Gênica
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